http://2012.igem.org/wiki/index.php?title=Special:Contributions&feed=atom&limit=500&target=Amurthur2012.igem.org - User contributions [en]2024-03-28T11:08:19ZFrom 2012.igem.orgMediaWiki 1.16.0http://2012.igem.org/Team:PennTeam:Penn2013-08-24T19:22:38Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<br />
<!-----------------------------------------------------------------------------------------------------><br />
<br />
<table width="1000px" height="335px" style="background:#01256e; margin-bottom:20px;"><br />
<tr style="text-align:center;" valign="top"><br />
<td style="width:750px;padding-top:20px;"><br />
<br />
<div class="rotator"><br />
<ul id="rotmenu"><br />
<li><br />
<a href="rot1" style="font-size:12px;"><b>Regional Winners!</b></a><br />
<div style="display:none;"><br />
<div class="info_image">2/2c/Regional-Winners-Slider.jpg</div><br />
<div class="info_heading"></div><br />
<div class="info_description"><br />
Penn iGEM placed first in the Americas East Regional Jamboree and advanced to the World Championships!<br />
<a href="https://igem.org/Results?year=2012&division=igem&region=Americas_East" class="more">Read more</a><br />
</div><br />
</div><br />
</li><br />
<li><br />
<a href="rot2" style="font-size:12px;"><p><b>Light Induced</b></p><p><b>Lysis</b></p></a><br />
<div style="display:none;"><br />
<div class="info_image">4/4f/Light-Induced-Lysis-Slider.jpg</div><br />
<div class="info_heading">pDAWN ClYA construct</div><br />
<br />
<div class="info_description">We have developed light-activated cell lysis using the YF1/FixJ Blue Light sensor and the ClyA protein.<br />
<br><br />
<a href="https://2012.igem.org/Team:Penn/DrugDeliveryResults" class="more">Read more</a><br />
</div><br />
</div><br />
</li><br />
<li><br />
<a href="rot3" style="font-size:12px;"><p><b>Novel Surface</b></p><p><b>Display</b></p></a><br />
<div style="display:none;"><br />
<div class="info_image">4/46/Novel-Surface-Display-Slider.jpg</div><br />
<div class="info_heading">Display of ANTi-her2 DARPin</div><br />
<div class="info_description"><br />
Our team is the first to use the INPNC protein to display the DARPin Anti-HER2 binding protein on the surface of bacteria <br><br />
<a href="https://2012.igem.org/Team:Penn/DrugDeliveryResults" class="more">Read more</a><br />
</div><br />
</div><br />
</li><br />
<li><br />
<a href="rot4" style="font-size:12px;"><b>Outreach</b></a><br />
<div style="display:none;"><br />
<div class="info_image">5/53/Outreach-Slider-Reverse.jpg</div><br />
<div class="info_heading">Clark Park Science Fair</div><br />
<div class="info_description"><br />
Learn more about our education outreach with high schoolers and West Philadelphia residents!<br />
<br><br />
<a href="https://2012.igem.org/Team:Penn/Outreach" class="more">Read more</a><br />
</div><br />
</div><br />
</li><br />
<li><br />
<a href="rot5" style="font-size:12px;"><b>Lab Work</b></a><br />
<div style="display:none;"><br />
<div class="info_image">3/36/Lab-Work-Slider2.jpg</div><br />
<div class="info_heading"></div><br />
<div class="info_description"><br />
Learn about cool experiments the team performed this summer!<br />
<br><br />
<a href="https://2012.igem.org/Team:Penn/Notebook" class="more">Read more</a><br />
</div><br />
</div><br />
</li><br />
<br />
</ul><br />
<div id="rot1"><br />
<img src="" width="720" height="300" class="bg" alt=""/><br />
<br />
<div class="description"><br />
<p></p><br />
<br />
</div> <br />
</div><br />
</div><br />
<br />
<!-- The JavaScript --><br />
<script type="text/javascript" src="http://ajax.googleapis.com/ajax/libs/jquery/1/jquery.min.js"></script><br />
<script type="text/javascript" src='http://cdnjs.cloudflare.com/ajax/libs/jquery-easing/1.3/jquery.easing.min.js?action=raw&amp;ctype=text/javascript'></script><br />
<br />
<script type="text/javascript"><br />
$(function() {<br />
var current = 1;<br />
<br />
var iterate = function(){<br />
var i = parseInt(current+1);<br />
var lis = $('#rotmenu').children('li').size();<br />
if(i>lis) i = 1;<br />
display($('#rotmenu li:nth-child('+i+')'));<br />
}<br />
display($('#rotmenu li:first'));<br />
var slidetime = setInterval(iterate,3800);<br />
<br />
$('#rotmenu li').bind('click',function(e){<br />
clearTimeout(slidetime);<br />
display($(this));<br />
e.preventDefault();<br />
});<br />
<br />
function display(elem){<br />
var $this = elem;<br />
var repeat = false;<br />
if(current == parseInt($this.index() + 1))<br />
repeat = true;<br />
<br />
if(!repeat)<br />
$this.parent().find('li:nth-child('+current+') a').stop(true,true).animate({'marginRight':'-20px'},300,function(){<br />
$(this).animate({'opacity':'0.7'},700);<br />
});<br />
<br />
current = parseInt($this.index() + 1);<br />
<br />
var elem = $('a',$this);<br />
<br />
elem.stop(true,true).animate({'marginRight':'0px','opacity':'1.0'},300);<br />
<br />
var info_elem = elem.next();<br />
<br />
<br />
<br />
$('#rot1 .description').animate({'bottom':'-270px'},500,'easeOutCirc',function(){<br />
$('p',$(this)).html(info_elem.find('.info_description').html());<br />
$(this).animate({'bottom':'0px'},400,'easeInOutQuad');<br />
})<br />
$('#rot1').prepend(<br />
$('<img/>',{<br />
style : 'opacity:0',<br />
className : 'bg'<br />
}).load(<br />
function(){<br />
$(this).animate({'opacity':'1'},600);<br />
$('#rot1 img:first').next().animate({'opacity':'0'},700,function(){<br />
$(this).remove();<br />
});<br />
}<br />
).attr('src','https://static.igem.org/mediawiki/2012/'+info_elem.find('.info_image').html()).attr('width','720').attr('height','300')<br />
);<br />
}<br />
});<br />
</script><br />
<!---------------------------------------- END slider ------------------------------------------------------------><br />
<br />
</td><br />
<td style="width:250px;"><br />
<table class="box" style="margin-top:20px;height:305px;width:220px;margin-left:10px;margin-right:10px;"><br />
<tr style="text-align:center;" valign="top"><br />
<td><br />
<div style="font-size:20px;height:200px;width:220px;"><br />
<div><br />
<h2><b>Welcome to the Penn Wiki!</b></h2><br />
<br />
<p style="text-align:center; font-size: 12px; font-weight:bold; font-color: white; padding-left:5px; padding-right:5px;"><br />
With the help of our sponsors, the Penn iGEM Team is excited to compete for its second year! </p><br />
<br />
<p style="text-align:center; font-size: 12px; font-weight:bold; font-color: white; padding-left:5px; padding-right:5px;"><br />
We have been working hard all summer and are proud of what we have accomplished. <br />
</p><br />
<br />
<p style="text-align:center; font-size: 12px; font-weight:bold; font-color: white; padding-left:5px; padding-right:5px;"><br />
Please, take a look around!<br />
</p><br />
<p style="text-align:center; font-size: 14px; font-weight:bold; font-color: white;"><u>Quick Links</u></p><br />
<br />
<br />
<p style="text-align:center; font-size: 14px; font-weight:bold; font-color: white;"><a href="https://2012.igem.org/Team:Penn/ProjectResults" class="white">Project Overview</a></p><br />
<br />
<br />
<p style="text-align:center; font-size: 14px; font-weight:bold; font-color: white;"><a href="https://2012.igem.org/Team:Penn/Parts" class="white">Parts Submitted</a></p><br />
<br />
<br />
<p style="text-align:center; font-size: 14px; font-weight:bold; font-color: white;"><a href="https://2012.igem.org/Team:Penn/Team" class="white">About the Team</a></p><br />
</div><br />
</div><br />
<br />
</td><br />
</tr><br />
</table><br />
<br />
</td><br />
</tr><br />
</table><br />
<br />
</td><br />
</tr><br />
</table><br />
<br />
<br />
<br />
<div style="background-color:#01256e;" align="center"><br />
<img src="https://static.igem.org/mediawiki/2012/7/7c/Accomplishments-Banner.gif" usemap="#banner" /><br />
<map id="banner"><br />
<area shape="rect" coords="51,4,293,295" href="https://2012.igem.org/Team:Penn/LightActivatedLysis" alt="" title="" /><br />
<area shape="rect" coords="380,4,622,295" href="https://2012.igem.org/Team:Penn/SurfaceDisplay" alt="" title="" /><br />
<area shape="rect" coords="707,4,949,295" href="https://2012.igem.org/Team:Penn/SurfaceDisplayBBa" alt="" title="" /><br />
<area shape="rect" coords="998,298,1000,300" href="http://www.image-maps.com/index.php?aff=mapped_users_4201210260637497" alt="Image Map" title="Image Map" /><br />
</map><br />
</div><br />
<br><br />
<br />
<div style="background-color:#01256e;" align="center"><br />
<iframe width="700" height="393" style="margin-left:10px;padding:10px; background-color:#000000;"src="http://www.youtube.com/embed/vadTEaV9Sk4" frameborder="1" allowfullscreen></iframe><br />
</div><br />
<br />
<br />
<table width="1000px" height="375px" style="margin-bottom:0px; margin-top:0px; padding-botttm:0px; background-color:#01256F;" ><br />
<tr style="text-align:center;" valign="top"><br />
<td style="width:650px; "><br />
<map id="sponsormap" name="sponsormap"><br />
<area shape="rect" coords="106,81,304,153" href="http://www.upenn.edu"><br />
<area shape="rect" coords="107,182,321,234" href="http://www.corning.com/lifesciences"><br />
<area shape="rect" coords="108,272,265,328" href="http://www.genewiz.com/"><br />
<area shape="rect" coords="284,260,387,337" href="http://www.upenn.edu/fisher/"><br />
<area shape="rect" coords="354,78,573,166" href="http://www.seas.upenn.edu/"><br />
<area shape="rect" coords="352,177,576,250" href="http://www.idtdna.com/site"><br />
<area shape="rect" coords="412,265,561,339" href="http://www.neb.com/nebecomm/default.asp"><br />
</map><br />
<img src="https://static.igem.org/mediawiki/2012/1/1d/NewSpons.jpg" width="650" height="400" usemap="#sponsormap"><br />
</td><br />
<br />
<td style="width:350px"><br />
<div style="margin-left:55px; padding-top:55px; padding-bottom:5px;"><br />
<br />
<script charset="utf-8" src="http://widgets.twimg.com/j/2/widget.js"></script><br />
<script><br />
new TWTR.Widget({<br />
version: 2,<br />
type: 'profile',<br />
rpp: 3,<br />
interval: 30000,<br />
width: 235,<br />
height: 185,<br />
theme: {<br />
shell: {<br />
background: '#01256e',<br />
color: '#ffffff'<br />
},<br />
tweets: {<br />
background: '#ffffff',<br />
<br />
color: '#000000',<br />
links: '#01256e'<br />
}<br />
},<br />
features: {<br />
scrollbar: true,<br />
loop: false,<br />
live: false,<br />
behavior: 'all'<br />
}<br />
}).render().setUser('PenniGEM').start();<br />
</script><br />
<br />
</div><br />
</td><br />
<br />
</tr><br />
<br />
</table><br><br />
<br />
</html></div>Amurthurhttp://2012.igem.org/Team:PennTeam:Penn2013-08-24T19:15:54Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<br />
<!-----------------------------------------------------------------------------------------------------><br />
<br />
<table width="1000px" height="335px" style="background:#01256e; margin-bottom:20px;"><br />
<tr style="text-align:center;" valign="top"><br />
<td style="width:750px;padding-top:20px;"><br />
<br />
<div class="rotator"><br />
<ul id="rotmenu"><br />
<li><br />
<a href="rot1" style="font-size:12px;"><b>Regional Winners!</b></a><br />
<div style="display:none;"><br />
<div class="info_image">2/2c/Regional-Winners-Slider.jpg</div><br />
<div class="info_heading"></div><br />
<div class="info_description"><br />
Penn iGEM placed first in the Americas East Regional Jamboree and advanced to the World Championships!<br />
<a href="https://igem.org/Results?year=2012&division=igem&region=Americas_East" class="more">Read more</a><br />
</div><br />
</div><br />
</li><br />
<li><br />
<a href="rot2" style="font-size:12px;"><p><b>Light Induced</b></p><p><b>Lysis</b></p></a><br />
<div style="display:none;"><br />
<div class="info_image">4/4f/Light-Induced-Lysis-Slider.jpg</div><br />
<div class="info_heading">pDAWN ClYA construct</div><br />
<br />
<div class="info_description">We have developed light-activated cell lysis using the YF1/FixJ Blue Light sensor and the ClyA protein.<br />
<br><br />
<a href="https://2012.igem.org/Team:Penn/DrugDeliveryResults" class="more">Read more</a><br />
</div><br />
</div><br />
</li><br />
<li><br />
<a href="rot3" style="font-size:12px;"><p><b>Novel Surface</b></p><p><b>Display</b></p></a><br />
<div style="display:none;"><br />
<div class="info_image">4/46/Novel-Surface-Display-Slider.jpg</div><br />
<div class="info_heading">Display of ANTi-her2 DARPin</div><br />
<div class="info_description"><br />
Our team is the first to use the INPNC protein to display the DARPin Anti-HER2 binding protein on the surface of bacteria <br><br />
<a href="https://2012.igem.org/Team:Penn/DrugDeliveryResults" class="more">Read more</a><br />
</div><br />
</div><br />
</li><br />
<li><br />
<a href="rot4" style="font-size:12px;"><b>Outreach</b></a><br />
<div style="display:none;"><br />
<div class="info_image">5/53/Outreach-Slider-Reverse.jpg</div><br />
<div class="info_heading">Clark Park Science Fair</div><br />
<div class="info_description"><br />
Learn more about our education outreach with high schoolers and West Philadelphia residents!<br />
<br><br />
<a href="https://2012.igem.org/Team:Penn/Outreach" class="more">Read more</a><br />
</div><br />
</div><br />
</li><br />
<li><br />
<a href="rot5" style="font-size:12px;"><b>Lab Work</b></a><br />
<div style="display:none;"><br />
<div class="info_image">3/36/Lab-Work-Slider2.jpg</div><br />
<div class="info_heading"></div><br />
<div class="info_description"><br />
Learn about cool experiments the team performed this summer!<br />
<br><br />
<a href="https://2012.igem.org/Team:Penn/Notebook" class="more">Read more</a><br />
</div><br />
</div><br />
</li><br />
<br />
</ul><br />
<div id="rot1"><br />
<img src="" width="720" height="300" class="bg" alt=""/><br />
<br />
<div class="description"><br />
<p></p><br />
<br />
</div> <br />
</div><br />
</div><br />
<br />
<!-- The JavaScript --><br />
<script type="text/javascript" src="http://ajax.googleapis.com/ajax/libs/jquery/1/jquery.min.js"></script><br />
<script type="text/javascript" src='http://cdnjs.cloudflare.com/ajax/libs/jquery-easing/1.3/jquery.easing.min.js?action=raw&amp;ctype=text/javascript'></script><br />
<br />
<script type="text/javascript"><br />
$(function() {<br />
var current = 1;<br />
<br />
var iterate = function(){<br />
var i = parseInt(current+1);<br />
var lis = $('#rotmenu').children('li').size();<br />
if(i>lis) i = 1;<br />
display($('#rotmenu li:nth-child('+i+')'));<br />
}<br />
display($('#rotmenu li:first'));<br />
var slidetime = setInterval(iterate,3800);<br />
<br />
$('#rotmenu li').bind('click',function(e){<br />
clearTimeout(slidetime);<br />
display($(this));<br />
e.preventDefault();<br />
});<br />
<br />
function display(elem){<br />
var $this = elem;<br />
var repeat = false;<br />
if(current == parseInt($this.index() + 1))<br />
repeat = true;<br />
<br />
if(!repeat)<br />
$this.parent().find('li:nth-child('+current+') a').stop(true,true).animate({'marginRight':'-20px'},300,function(){<br />
$(this).animate({'opacity':'0.7'},700);<br />
});<br />
<br />
current = parseInt($this.index() + 1);<br />
<br />
var elem = $('a',$this);<br />
<br />
elem.stop(true,true).animate({'marginRight':'0px','opacity':'1.0'},300);<br />
<br />
var info_elem = elem.next();<br />
<br />
<br />
<br />
$('#rot1 .description').animate({'bottom':'-270px'},500,'easeOutCirc',function(){<br />
$('p',$(this)).html(info_elem.find('.info_description').html());<br />
$(this).animate({'bottom':'0px'},400,'easeInOutQuad');<br />
})<br />
$('#rot1').prepend(<br />
$('<img/>',{<br />
style : 'opacity:0',<br />
className : 'bg'<br />
}).load(<br />
function(){<br />
$(this).animate({'opacity':'1'},600);<br />
$('#rot1 img:first').next().animate({'opacity':'0'},700,function(){<br />
$(this).remove();<br />
});<br />
}<br />
).attr('src','https://static.igem.org/mediawiki/2012/'+info_elem.find('.info_image').html()).attr('width','720').attr('height','300')<br />
);<br />
}<br />
});<br />
</script><br />
<!---------------------------------------- END slider ------------------------------------------------------------><br />
<br />
</td><br />
<td style="width:250px;"><br />
<table class="box" style="margin-top:20px;height:305px;width:220px;margin-left:10px;margin-right:10px;"><br />
<tr style="text-align:center;" valign="top"><br />
<td><br />
<div style="font-size:20px;height:200px;width:220px;"><br />
<div><br />
<h2><b>Welcome to the Penn Wiki!</b></h2><br />
<br />
<p style="text-align:center; font-size: 12px; font-weight:bold; font-color: white; padding-left:5px; padding-right:5px;"><br />
With the help of our sponsors, the Penn iGEM Team is excited to compete for its second year! </p><br />
<br />
<p style="text-align:center; font-size: 12px; font-weight:bold; font-color: white; padding-left:5px; padding-right:5px;"><br />
We have been working hard all summer and are proud of what we have accomplished. <br />
</p><br />
<br />
<p style="text-align:center; font-size: 12px; font-weight:bold; font-color: white; padding-left:5px; padding-right:5px;"><br />
Please, take a look around!<br />
</p><br />
<p style="text-align:center; font-size: 14px; font-weight:bold; font-color: white;"><u>Quick Links</u></p><br />
<br />
<br />
<p style="text-align:center; font-size: 14px; font-weight:bold; font-color: white;"><a href="https://2012.igem.org/Team:Penn/ProjectResults" class="white">Project Overview</a></p><br />
<br />
<br />
<p style="text-align:center; font-size: 14px; font-weight:bold; font-color: white;"><a href="https://2012.igem.org/Team:Penn/Parts" class="white">Parts Submitted</a></p><br />
<br />
<br />
<p style="text-align:center; font-size: 14px; font-weight:bold; font-color: white;"><a href="https://2012.igem.org/Team:Penn/Team" class="white">About the Team</a></p><br />
</div><br />
</div><br />
<br />
</td><br />
</tr><br />
</table><br />
<br />
</td><br />
</tr><br />
</table><br />
<br />
</td><br />
</tr><br />
</table><br />
<br />
<br />
<br />
<div style="background-color:#01256e;" align="center"><br />
<img src="https://static.igem.org/mediawiki/2012/7/7c/Accomplishments-Banner.gif" usemap="#banner" /><br />
<map id="banner"><br />
<area shape="rect" coords="51,4,293,295" href="https://2012.igem.org/Team:Penn/LightActivatedLysis" alt="" title="" /><br />
<area shape="rect" coords="380,4,622,295" href="https://2012.igem.org/Team:Penn/SurfaceDisplay" alt="" title="" /><br />
<area shape="rect" coords="707,4,949,295" href="https://2012.igem.org/Team:Penn/SurfaceDisplayBBa" alt="" title="" /><br />
<area shape="rect" coords="998,298,1000,300" href="http://www.image-maps.com/index.php?aff=mapped_users_4201210260637497" alt="Image Map" title="Image Map" /><br />
</map><br />
</div><br />
<br><br />
<br />
<div style="background-color:#01256e;" align="center"><br />
<iframe width="700" height="393" style="margin-left:10px;padding:10px; background-color:#000000;"src="http://www.youtube.com/embed/vadTEaV9Sk4" frameborder="1" allowfullscreen></iframe><br />
</div><br />
<br />
<br />
<table width="1000px" height="375px" style="margin-bottom:0px; margin-top:0px; padding-botttm:0px; background-color:#01256F;" ><br />
<tr style="text-align:center;" valign="top"><br />
<td style="width:650px; "><br />
<map id="sponsormap" name="sponsormap"><br />
<area shape="rect" coords="106,81,304,153" href="http://www.upenn.edu"><br />
<area shape="rect" coords="107,182,321,234" href="http://www.corning.com/lifesciences"><br />
<area shape="rect" coords="108,272,265,328" href="http://www.genewiz.com/"><br />
<area shape="rect" coords="284,260,387,337" href="http://www.upenn.edu/fisher/"><br />
<area shape="rect" coords="354,78,573,166" href="http://www.seas.upenn.edu/"><br />
<area shape="rect" coords="352,177,576,250" href="http://www.idtdna.com/site"><br />
<area shape="rect" coords="412,265,561,339" href="http://www.neb.com/nebecomm/default.asp"><br />
</map><br />
<img src="https://static.igem.org/mediawiki/2012/1/1d/NewSpons.jpg" width="650" height="400" usemap="#sponsormap"><br />
</td><br />
<br />
<td style="width:350px"><br />
<div style="margin-left:55px; padding-top:55px; padding-bottom:5px;"><br />
<!-- <br />
<script charset="utf-8" src="http://widgets.twimg.com/j/2/widget.js"></script><br />
<script><br />
new TWTR.Widget({<br />
version: 2,<br />
type: 'profile',<br />
rpp: 3,<br />
interval: 30000,<br />
width: 235,<br />
height: 185,<br />
theme: {<br />
shell: {<br />
background: '#01256e',<br />
color: '#ffffff'<br />
},<br />
tweets: {<br />
background: '#ffffff',<br />
<br />
color: '#000000',<br />
links: '#01256e'<br />
}<br />
},<br />
features: {<br />
scrollbar: true,<br />
loop: false,<br />
live: false,<br />
behavior: 'all'<br />
}<br />
}).render().setUser('PenniGEM').start();<br />
</script><br />
--><br />
</div><br />
</td><br />
<br />
</tr><br />
<br />
</table><br><br />
<br />
</html></div>Amurthurhttp://2012.igem.org/Team:PennTeam:Penn2013-08-24T19:09:36Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<br />
<!-----------------------------------------------------------------------------------------------------><br />
<br />
<table width="1000px" height="335px" style="background:#01256e; margin-bottom:20px;"><br />
<tr style="text-align:center;" valign="top"><br />
<td style="width:750px;padding-top:20px;"><br />
<br />
<div class="rotator"><br />
<ul id="rotmenu"><br />
<li><br />
<a href="rot1" style="font-size:12px;"><b>Regional Winners!</b></a><br />
<div style="display:none;"><br />
<div class="info_image">2/2c/Regional-Winners-Slider.jpg</div><br />
<div class="info_heading"></div><br />
<div class="info_description"><br />
Penn iGEM placed first in the Americas East Regional Jamboree and advanced to the World Championships!<br />
<a href="https://igem.org/Results?year=2012&division=igem&region=Americas_East" class="more">Read more</a><br />
</div><br />
</div><br />
</li><br />
<li><br />
<a href="rot2" style="font-size:12px;"><p><b>Light Induced</b></p><p><b>Lysis</b></p></a><br />
<div style="display:none;"><br />
<div class="info_image">4/4f/Light-Induced-Lysis-Slider.jpg</div><br />
<div class="info_heading">pDAWN ClYA construct</div><br />
<br />
<div class="info_description">We have developed light-activated cell lysis using the YF1/FixJ Blue Light sensor and the ClyA protein.<br />
<br><br />
<a href="https://2012.igem.org/Team:Penn/DrugDeliveryResults" class="more">Read more</a><br />
</div><br />
</div><br />
</li><br />
<li><br />
<a href="rot3" style="font-size:12px;"><p><b>Novel Surface</b></p><p><b>Display</b></p></a><br />
<div style="display:none;"><br />
<div class="info_image">4/46/Novel-Surface-Display-Slider.jpg</div><br />
<div class="info_heading">Display of ANTi-her2 DARPin</div><br />
<div class="info_description"><br />
Our team is the first to use the INPNC protein to display the DARPin Anti-HER2 binding protein on the surface of bacteria <br><br />
<a href="https://2012.igem.org/Team:Penn/DrugDeliveryResults" class="more">Read more</a><br />
</div><br />
</div><br />
</li><br />
<li><br />
<a href="rot4" style="font-size:12px;"><b>Outreach</b></a><br />
<div style="display:none;"><br />
<div class="info_image">5/53/Outreach-Slider-Reverse.jpg</div><br />
<div class="info_heading">Clark Park Science Fair</div><br />
<div class="info_description"><br />
Learn more about our education outreach with high schoolers and West Philadelphia residents!<br />
<br><br />
<a href="https://2012.igem.org/Team:Penn/Outreach" class="more">Read more</a><br />
</div><br />
</div><br />
</li><br />
<li><br />
<a href="rot5" style="font-size:12px;"><b>Lab Work</b></a><br />
<div style="display:none;"><br />
<div class="info_image">3/36/Lab-Work-Slider2.jpg</div><br />
<div class="info_heading"></div><br />
<div class="info_description"><br />
Learn about cool experiments the team performed this summer!<br />
<br><br />
<a href="https://2012.igem.org/Team:Penn/Notebook" class="more">Read more</a><br />
</div><br />
</div><br />
</li><br />
<br />
</ul><br />
<div id="rot1"><br />
<img src="" width="720" height="300" class="bg" alt=""/><br />
<br />
<div class="description"><br />
<p></p><br />
<br />
</div> <br />
</div><br />
</div><br />
<br />
<!-- The JavaScript --><br />
<script type="text/javascript" src="http://ajax.googleapis.com/ajax/libs/jquery/1/jquery.min.js"></script><br />
<script type="text/javascript" src='http://cdnjs.cloudflare.com/ajax/libs/jquery-easing/1.3/jquery.easing.min.js?action=raw&amp;ctype=text/javascript'></script><br />
<br />
<script type="text/javascript"><br />
$(function() {<br />
var current = 1;<br />
<br />
var iterate = function(){<br />
var i = parseInt(current+1);<br />
var lis = $('#rotmenu').children('li').size();<br />
if(i>lis) i = 1;<br />
display($('#rotmenu li:nth-child('+i+')'));<br />
}<br />
display($('#rotmenu li:first'));<br />
var slidetime = setInterval(iterate,3800);<br />
<br />
$('#rotmenu li').bind('click',function(e){<br />
clearTimeout(slidetime);<br />
display($(this));<br />
e.preventDefault();<br />
});<br />
<br />
function display(elem){<br />
var $this = elem;<br />
var repeat = false;<br />
if(current == parseInt($this.index() + 1))<br />
repeat = true;<br />
<br />
if(!repeat)<br />
$this.parent().find('li:nth-child('+current+') a').stop(true,true).animate({'marginRight':'-20px'},300,function(){<br />
$(this).animate({'opacity':'0.7'},700);<br />
});<br />
<br />
current = parseInt($this.index() + 1);<br />
<br />
var elem = $('a',$this);<br />
<br />
elem.stop(true,true).animate({'marginRight':'0px','opacity':'1.0'},300);<br />
<br />
var info_elem = elem.next();<br />
<br />
<br />
<br />
$('#rot1 .description').animate({'bottom':'-270px'},500,'easeOutCirc',function(){<br />
$('p',$(this)).html(info_elem.find('.info_description').html());<br />
$(this).animate({'bottom':'0px'},400,'easeInOutQuad');<br />
})<br />
$('#rot1').prepend(<br />
$('<img/>',{<br />
style : 'opacity:0',<br />
className : 'bg'<br />
}).load(<br />
function(){<br />
$(this).animate({'opacity':'1'},600);<br />
$('#rot1 img:first').next().animate({'opacity':'0'},700,function(){<br />
$(this).remove();<br />
});<br />
}<br />
).attr('src','https://static.igem.org/mediawiki/2012/'+info_elem.find('.info_image').html()).attr('width','720').attr('height','300')<br />
);<br />
}<br />
});<br />
</script><br />
<!---------------------------------------- END slider ------------------------------------------------------------><br />
<br />
</td><br />
<td style="width:250px;"><br />
<table class="box" style="margin-top:20px;height:305px;width:220px;margin-left:10px;margin-right:10px;"><br />
<tr style="text-align:center;" valign="top"><br />
<td><br />
<div style="font-size:20px;height:200px;width:220px;"><br />
<div><br />
<h2><b>Welcome to the Penn Wiki!</b></h2><br />
<br />
<p style="text-align:center; font-size: 12px; font-weight:bold; font-color: white; padding-left:5px; padding-right:5px;"><br />
With the help of our sponsors, the Penn iGEM Team is excited to compete for its second year! </p><br />
<br />
<p style="text-align:center; font-size: 12px; font-weight:bold; font-color: white; padding-left:5px; padding-right:5px;"><br />
We have been working hard all summer and are proud of what we have accomplished. <br />
</p><br />
<br />
<p style="text-align:center; font-size: 12px; font-weight:bold; font-color: white; padding-left:5px; padding-right:5px;"><br />
Please, take a look around!<br />
</p><br />
<p style="text-align:center; font-size: 14px; font-weight:bold; font-color: white;"><u>Quick Links</u></p><br />
<br />
<br />
<p style="text-align:center; font-size: 14px; font-weight:bold; font-color: white;"><a href="https://2012.igem.org/Team:Penn/ProjectResults" class="white">Project Overview</a></p><br />
<br />
<br />
<p style="text-align:center; font-size: 14px; font-weight:bold; font-color: white;"><a href="https://2012.igem.org/Team:Penn/Parts" class="white">Parts Submitted</a></p><br />
<br />
<br />
<p style="text-align:center; font-size: 14px; font-weight:bold; font-color: white;"><a href="https://2012.igem.org/Team:Penn/Team" class="white">About the Team</a></p><br />
</div><br />
</div><br />
<br />
</td><br />
</tr><br />
</table><br />
<br />
</td><br />
</tr><br />
</table><br />
<br />
</td><br />
</tr><br />
</table><br />
<br />
<br />
<br />
<div style="background-color:#01256e;" align="center"><br />
<img src="https://static.igem.org/mediawiki/2012/7/7c/Accomplishments-Banner.gif" usemap="#banner" /><br />
<map id="banner"><br />
<area shape="rect" coords="51,4,293,295" href="https://2012.igem.org/Team:Penn/LightActivatedLysis" alt="" title="" /><br />
<area shape="rect" coords="380,4,622,295" href="https://2012.igem.org/Team:Penn/SurfaceDisplay" alt="" title="" /><br />
<area shape="rect" coords="707,4,949,295" href="https://2012.igem.org/Team:Penn/SurfaceDisplayBBa" alt="" title="" /><br />
<area shape="rect" coords="998,298,1000,300" href="http://www.image-maps.com/index.php?aff=mapped_users_4201210260637497" alt="Image Map" title="Image Map" /><br />
</map><br />
</div><br />
<br><br />
<br />
<div style="background-color:#01256e;" align="center"><br />
<iframe width="700" height="393" style="margin-left:10px;padding:10px; background-color:#000000;"src="http://www.youtube.com/embed/vadTEaV9Sk4" frameborder="1" allowfullscreen></iframe><br />
</div><br />
<br />
<br />
<table width="1000px" height="375px" style="margin-bottom:0px; margin-top:0px; padding-botttm:0px; background-color:#01256F;" ><br />
<tr style="text-align:center;" valign="top"><br />
<td style="width:650px; "><br />
<map id="sponsormap" name="sponsormap"><br />
<area shape="rect" coords="106,81,304,153" href="http://www.upenn.edu"><br />
<area shape="rect" coords="107,182,321,234" href="http://www.corning.com/lifesciences"><br />
<area shape="rect" coords="108,272,265,328" href="http://www.genewiz.com/"><br />
<area shape="rect" coords="284,260,387,337" href="http://www.upenn.edu/fisher/"><br />
<area shape="rect" coords="354,78,573,166" href="http://www.seas.upenn.edu/"><br />
<area shape="rect" coords="352,177,576,250" href="http://www.idtdna.com/site"><br />
<area shape="rect" coords="412,265,561,339" href="http://www.neb.com/nebecomm/default.asp"><br />
</map><br />
<img src="https://static.igem.org/mediawiki/2012/1/1d/NewSpons.jpg" width="650" height="400" usemap="#sponsormap"><br />
</td><br />
<br />
<td style="width:350px"><br />
<div style="margin-left:55px; padding-top:55px; padding-bottom:5px;"><br />
<script charset="utf-8" src="http://widgets.twimg.com/j/2/widget.js"></script><br />
<script><br />
new TWTR.Widget({<br />
version: 2,<br />
type: 'profile',<br />
rpp: 3,<br />
interval: 30000,<br />
width: 235,<br />
height: 185,<br />
theme: {<br />
shell: {<br />
background: '#01256e',<br />
color: '#ffffff'<br />
},<br />
tweets: {<br />
background: '#ffffff',<br />
<br />
color: '#000000',<br />
links: '#01256e'<br />
}<br />
},<br />
features: {<br />
scrollbar: true,<br />
loop: false,<br />
live: false,<br />
behavior: 'all'<br />
}<br />
}).render().setUser('PenniGEM').start();<br />
</script><br />
</div><br />
</td><br />
<br />
</tr><br />
<br />
</table><br><br />
<br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/Template/SiteTeam:Penn/Template/Site2013-08-24T19:08:24Z<p>Amurthur: </p>
<hr />
<div><html><br />
<head><br />
<!-- coded by Ashwin Amurthur --><br />
<!-- University of Pennsylvania iGEM team WIKI --><br />
<link rel="stylesheet" href='/Team:Penn/css/navigation?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/layerslider?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/sliderstyle?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/style2?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/columns?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/igemcss?action=raw&amp;ctype=text/css' /><br />
<script type="text/javascript" src='/Team:Penn/js/accordion?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/jquery-1.7.1.min.js?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/layersliderkreatura?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/cufon-replace.js?action=raw&amp;ctype=text/javascript'></script><br />
<br />
<title>Penn 2012 iGEM Wiki</title><br />
<br />
<style type="text/css"><br />
<br />
a.white:link {color:#FFF;} /* unvisited link */<br />
a.white:visited {color:#FFF;} /* visited link */<br />
a.white:hover {color:#FF;} /* mouse over link */<br />
a.white:active {color:#FFF;} /* selected link */ <br />
<br />
<br />
#top-section2 {<br />
width:1000px;<br />
border:0px;<br />
height:0px;<br />
padding-bottom:0px;<br />
background: #01256e;<br />
margin-bottom: 0px;<br />
margin-top: 0px;<br />
}<br />
<br />
<br />
</style><br />
<br />
</head><br />
<br />
<body><br />
<div id="top-section"><br />
<div id="p-logo2"><br />
<a href="https://2012.igem.org/Team:Penn"><img src='https://static.igem.org/mediawiki/2012/f/fa/Sponsor-Image-Wiki-Blue-Background_v2.gif' width="1000" height="175" usemap="#igemmap"></a><br />
<br />
<br />
<map name="igemmap" ><br />
<area shape="rect" coords="0,122,67,170" href="https://2012.igem.org" alt="" title="" /><br />
<area shape="rect" coords="913,131,995,170" href="http://www.upenn.edu" alt="" title="" /><br />
<area shape="rect" coords="998,173,1000,175" href="http://www.image-maps.com/index.php?aff=mapped_users_1201210022332192" alt="Image Map" title="Image Map" /><br />
</map><br />
<br />
<br />
</div><br />
<br />
</div><br />
</div><br />
<br />
<div style="margin-top:0px;margin-bottom:0px; padding-bottom:0px;"><br />
<a href="https://2012.igem.org/Team:Penn" style="padding:0px; margin:0px;"><br />
<img src="https://static.igem.org/mediawiki/2012/1/10/Bgblue.png" /></a><br />
</div><br />
<br />
<br />
<ul class="nav"><br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/ProjectOverview" style="font-size:13px;">Our Project</a><br />
<br />
<ul><br />
<li><a href='/Team:Penn/ProjectOverview'>Motivation</a></li><br />
<li><a href='/Team:Penn/ProjectResults'>Our Therapeutic</a></li><br />
<li><a href='/Team:Penn/Achievements'>Achievements</a></li><br />
<br />
<br />
<br />
</ul><br />
</li> <br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/LightActivatedOverview" style="font-size:13px;">Light Activation</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/LightActivatedOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/BLSensor'>YF1/FixJ BL Sensor</a></li><br />
<li><a href='/Team:Penn/LightActivatedLysis'>Light Activated Cell Lysis</a></li><br />
<br />
<br />
</ul><br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/SurfaceDisplayOverview" style="font-size:13px;">Surface Display</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/SurfaceDisplayOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/SurfaceDisplay'>INPNC Surface Display</a></li><br />
<li><a href='/Team:Penn/Targeting'>HER2 Targeting </a></li><br />
<li><a href='/Team:Penn/SurfaceDisplayBBa'>BBa Surface Display Platform</a></li><br />
<br />
</ul><br />
<br />
</li> <br />
<br />
<br />
<li class="dropdown"><br />
<br />
<a href="#" style="font-size:13px;">Wet Lab</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/Notebook'>Notebook</a></li><br />
<li><a href='/Team:Penn/Safety'>Safety</a></li><br />
<li><a href='/Team:Penn/Protocols'>Protocols</a></li><br />
<li><a href='/Team:Penn/Parts'>Parts</a></li><br />
<br />
</ul><br />
<br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/HumanPracticesOverview" style="font-size:13px;">Human Practices</a><br />
<br />
<ul><br />
<li><a href='/Team:Penn/HumanPracticesOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/Barriers'>Bacterial Therapeutics</a></li> <br />
<li><a href='/Team:Penn/Outreach'>Outreach</a></li><br />
<li><a href='/Team:Penn/Perceptions'>Public Perception</a></li><br />
<li><a href='/Team:Penn/Nissle'>E. Coli Nissle 1917</a></li><br />
<li><a href='/Team:Penn/VerifiGEM'>VerifiGEM</a></li><br />
</ul> <br />
<br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/Team" style="font-size:13px;">About Us</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/Team'>Team</a></li> <br />
<li><a href='/Team:Penn/Sponsors'>Sponsors</a></li><br />
</ul> <br />
</li><br />
<br />
<div class="arrow"></div><br />
<br />
</ul><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/Template/SiteTeam:Penn/Template/Site2013-08-24T19:03:27Z<p>Amurthur: </p>
<hr />
<div><html><br />
<head><br />
<!-- coded by Ashwin Amurthur --><br />
<!-- University of Pennsylvania iGEM team WIKI --><br />
<link rel="stylesheet" href='/Team:Penn/css/navigation?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/layerslider?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/sliderstyle?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/style2?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/columns?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/igemcss?action=raw&amp;ctype=text/css' /><br />
<script type="text/javascript" src='/Team:Penn/js/accordion?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/jquery-1.7.1.min.js?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='http://cdnjs.cloudflare.com/ajax/libs/jquery-easing/1.3/jquery.easing.mins.js'></script><br />
<script type="text/javascript" src='/Team:Penn/js/layersliderkreatura?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/cufon-replace.js?action=raw&amp;ctype=text/javascript'></script><br />
<br />
<title>Penn 2012 iGEM Wiki</title><br />
<br />
<style type="text/css"><br />
<br />
a.white:link {color:#FFF;} /* unvisited link */<br />
a.white:visited {color:#FFF;} /* visited link */<br />
a.white:hover {color:#FF;} /* mouse over link */<br />
a.white:active {color:#FFF;} /* selected link */ <br />
<br />
<br />
#top-section2 {<br />
width:1000px;<br />
border:0px;<br />
height:0px;<br />
padding-bottom:0px;<br />
background: #01256e;<br />
margin-bottom: 0px;<br />
margin-top: 0px;<br />
}<br />
<br />
<br />
</style><br />
<br />
</head><br />
<br />
<body><br />
<div id="top-section"><br />
<div id="p-logo2"><br />
<a href="https://2012.igem.org/Team:Penn"><img src='https://static.igem.org/mediawiki/2012/f/fa/Sponsor-Image-Wiki-Blue-Background_v2.gif' width="1000" height="175" usemap="#igemmap"></a><br />
<br />
<br />
<map name="igemmap" ><br />
<area shape="rect" coords="0,122,67,170" href="https://2012.igem.org" alt="" title="" /><br />
<area shape="rect" coords="913,131,995,170" href="http://www.upenn.edu" alt="" title="" /><br />
<area shape="rect" coords="998,173,1000,175" href="http://www.image-maps.com/index.php?aff=mapped_users_1201210022332192" alt="Image Map" title="Image Map" /><br />
</map><br />
<br />
<br />
</div><br />
<br />
</div><br />
</div><br />
<br />
<div style="margin-top:0px;margin-bottom:0px; padding-bottom:0px;"><br />
<a href="https://2012.igem.org/Team:Penn" style="padding:0px; margin:0px;"><br />
<img src="https://static.igem.org/mediawiki/2012/1/10/Bgblue.png" /></a><br />
</div><br />
<br />
<br />
<ul class="nav"><br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/ProjectOverview" style="font-size:13px;">Our Project</a><br />
<br />
<ul><br />
<li><a href='/Team:Penn/ProjectOverview'>Motivation</a></li><br />
<li><a href='/Team:Penn/ProjectResults'>Our Therapeutic</a></li><br />
<li><a href='/Team:Penn/Achievements'>Achievements</a></li><br />
<br />
<br />
<br />
</ul><br />
</li> <br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/LightActivatedOverview" style="font-size:13px;">Light Activation</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/LightActivatedOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/BLSensor'>YF1/FixJ BL Sensor</a></li><br />
<li><a href='/Team:Penn/LightActivatedLysis'>Light Activated Cell Lysis</a></li><br />
<br />
<br />
</ul><br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/SurfaceDisplayOverview" style="font-size:13px;">Surface Display</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/SurfaceDisplayOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/SurfaceDisplay'>INPNC Surface Display</a></li><br />
<li><a href='/Team:Penn/Targeting'>HER2 Targeting </a></li><br />
<li><a href='/Team:Penn/SurfaceDisplayBBa'>BBa Surface Display Platform</a></li><br />
<br />
</ul><br />
<br />
</li> <br />
<br />
<br />
<li class="dropdown"><br />
<br />
<a href="#" style="font-size:13px;">Wet Lab</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/Notebook'>Notebook</a></li><br />
<li><a href='/Team:Penn/Safety'>Safety</a></li><br />
<li><a href='/Team:Penn/Protocols'>Protocols</a></li><br />
<li><a href='/Team:Penn/Parts'>Parts</a></li><br />
<br />
</ul><br />
<br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/HumanPracticesOverview" style="font-size:13px;">Human Practices</a><br />
<br />
<ul><br />
<li><a href='/Team:Penn/HumanPracticesOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/Barriers'>Bacterial Therapeutics</a></li> <br />
<li><a href='/Team:Penn/Outreach'>Outreach</a></li><br />
<li><a href='/Team:Penn/Perceptions'>Public Perception</a></li><br />
<li><a href='/Team:Penn/Nissle'>E. Coli Nissle 1917</a></li><br />
<li><a href='/Team:Penn/VerifiGEM'>VerifiGEM</a></li><br />
</ul> <br />
<br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/Team" style="font-size:13px;">About Us</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/Team'>Team</a></li> <br />
<li><a href='/Team:Penn/Sponsors'>Sponsors</a></li><br />
</ul> <br />
</li><br />
<br />
<div class="arrow"></div><br />
<br />
</ul><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/Template/Site2Team:Penn/Template/Site22012-10-27T04:05:13Z<p>Amurthur: </p>
<hr />
<div><html><br />
<head><br />
<!-- coded by Ashwin Amurthur --><br />
<!-- University of Pennsylvania iGEM team WIKI --><br />
<link rel="stylesheet" href='/Team:Penn/css/navigation?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/layerslider?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/sliderstyle?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/style2?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/columns?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/igemcss?action=raw&amp;ctype=text/css' /><br />
<script type="text/javascript" src='/Team:Penn/js/accordion?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/jquery-1.7.1.min.js?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/layersliderkreatura?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/cufon-replace.js?action=raw&amp;ctype=text/javascript'></script><br />
<br />
<title>Penn 2012 iGEM Wiki</title><br />
<br />
<style type="text/css"><br />
<br />
a.white:link {color:#FFF;} /* unvisited link */<br />
a.white:visited {color:#FFF;} /* visited link */<br />
a.white:hover {color:#FF;} /* mouse over link */<br />
a.white:active {color:#FFF;} /* selected link */ <br />
<br />
<br />
#top-section2 {<br />
width:1000px;<br />
border:0px;<br />
height:0px;<br />
padding-bottom:0px;<br />
background: #01256e;<br />
margin-bottom: 0px;<br />
margin-top: 0px;<br />
}<br />
<br />
<br />
</style><br />
<br />
</head><br />
<br />
<body><br />
<div id="top-section"><br />
<div id="p-logo2"><br />
<a href="https://2012.igem.org/Team:Penn"><img src='https://static.igem.org/mediawiki/2012/f/fa/Sponsor-Image-Wiki-Blue-Background_v2.gif' width="1000" height="175" usemap="#igemmap"></a><br />
<br />
<br />
<map name="igemmap" ><br />
<area shape="rect" coords="0,122,67,170" href="https://2012.igem.org" alt="" title="" /><br />
<area shape="rect" coords="913,131,995,170" href="http://www.upenn.edu" alt="" title="" /><br />
<area shape="rect" coords="998,173,1000,175" href="http://www.image-maps.com/index.php?aff=mapped_users_1201210022332192" alt="Image Map" title="Image Map" /><br />
</map><br />
<br />
<br />
</div><br />
<br />
</div><br />
</div><br />
<br />
<div style="margin-top:0px;margin-bottom:0px; padding-bottom:0px;"><br />
<a href="https://2012.igem.org/Team:Penn" style="padding:0px; margin:0px;"><br />
<img src="https://static.igem.org/mediawiki/2012/1/10/Bgblue.png" /></a><br />
</div><br />
<br />
<br />
<ul class="nav"><br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/ProjectOverview" style="font-size:13px;">Our Project</a><br />
<br />
<ul><br />
<li><a href='/Team:Penn/ProjectOverview'>Motivation</a></li><br />
<li><a href='/Team:Penn/ProjectResults'>Our Therapeutic</a></li><br />
<li><a href='/Team:Penn/Achievements'>Achievements</a></li><br />
<br />
<br />
<br />
</ul><br />
</li> <br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/LightActivatedOverview" style="font-size:13px;">Light Activation</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/LightActivatedOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/BLSensor'>YF1/FixJ BL Sensor</a></li><br />
<li><a href='/Team:Penn/LightActivatedLysis'>Light Activated Cell Lysis</a></li><br />
<br />
<br />
</ul><br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/SurfaceDisplayOverview" style="font-size:13px;">Surface Display</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/SurfaceDisplayOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/SurfaceDisplay'>INPNC Surface Display</a></li><br />
<li><a href='/Team:Penn/Targeting'>HER2 Targeting </a></li><br />
<li><a href='/Team:Penn/SurfaceDisplayBBa'>BBa Surface Display Platform</a></li><br />
<br />
</ul><br />
<br />
</li> <br />
<br />
<br />
<li class="dropdown"><br />
<br />
<a href="#" style="font-size:13px;">Wet Lab</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/Notebook'>Notebook</a></li><br />
<li><a href='/Team:Penn/Safety'>Safety</a></li><br />
<li><a href='/Team:Penn/Protocols'>Protocols</a></li><br />
<li><a href='/Team:Penn/Parts'>Parts</a></li><br />
<br />
</ul><br />
<br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/HumanPracticesOverview" style="font-size:13px;">Human Practices</a><br />
<br />
<ul><br />
<li><a href='/Team:Penn/HumanPracticesOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/Barriers'>Bacterial Therapeutics</a></li> <br />
<li><a href='/Team:Penn/Outreach'>Outreach</a></li><br />
<li><a href='/Team:Penn/Perceptions'>Public Perception</a></li><br />
<li><a href='/Team:Penn/Nissle'>E. Coli Nissle 1917</a></li><br />
<li><a href='/Team:Penn/VerifiGEM'>VerifiGEM</a></li><br />
</ul> <br />
<br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/Team" style="font-size:13px;">About Us</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/Team'>Team</a></li> <br />
<li><a href='/Team:Penn/Sponsors'>Sponsors</a></li><br />
</ul> <br />
</li><br />
<br />
<div class="arrow"></div><br />
<br />
</ul><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/Template/SiteTeam:Penn/Template/Site2012-10-27T04:05:00Z<p>Amurthur: </p>
<hr />
<div><html><br />
<head><br />
<!-- coded by Ashwin Amurthur --><br />
<!-- University of Pennsylvania iGEM team WIKI --><br />
<link rel="stylesheet" href='/Team:Penn/css/navigation?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/layerslider?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/sliderstyle?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/style2?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/columns?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/igemcss?action=raw&amp;ctype=text/css' /><br />
<script type="text/javascript" src='/Team:Penn/js/accordion?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/jquery-1.7.1.min.js?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/layersliderkreatura?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/cufon-replace.js?action=raw&amp;ctype=text/javascript'></script><br />
<br />
<title>Penn 2012 iGEM Wiki</title><br />
<br />
<style type="text/css"><br />
<br />
a.white:link {color:#FFF;} /* unvisited link */<br />
a.white:visited {color:#FFF;} /* visited link */<br />
a.white:hover {color:#FF;} /* mouse over link */<br />
a.white:active {color:#FFF;} /* selected link */ <br />
<br />
<br />
#top-section2 {<br />
width:1000px;<br />
border:0px;<br />
height:0px;<br />
padding-bottom:0px;<br />
background: #01256e;<br />
margin-bottom: 0px;<br />
margin-top: 0px;<br />
}<br />
<br />
<br />
</style><br />
<br />
</head><br />
<br />
<body><br />
<div id="top-section"><br />
<div id="p-logo2"><br />
<a href="https://2012.igem.org/Team:Penn"><img src='https://static.igem.org/mediawiki/2012/f/fa/Sponsor-Image-Wiki-Blue-Background_v2.gif' width="1000" height="175" usemap="#igemmap"></a><br />
<br />
<br />
<map name="igemmap" ><br />
<area shape="rect" coords="0,122,67,170" href="https://2012.igem.org" alt="" title="" /><br />
<area shape="rect" coords="913,131,995,170" href="http://www.upenn.edu" alt="" title="" /><br />
<area shape="rect" coords="998,173,1000,175" href="http://www.image-maps.com/index.php?aff=mapped_users_1201210022332192" alt="Image Map" title="Image Map" /><br />
</map><br />
<br />
<br />
</div><br />
<br />
</div><br />
</div><br />
<br />
<div style="margin-top:0px;margin-bottom:0px; padding-bottom:0px;"><br />
<a href="https://2012.igem.org/Team:Penn" style="padding:0px; margin:0px;"><br />
<img src="https://static.igem.org/mediawiki/2012/1/10/Bgblue.png" /></a><br />
</div><br />
<br />
<br />
<ul class="nav"><br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/ProjectOverview" style="font-size:13px;">Our Project</a><br />
<br />
<ul><br />
<li><a href='/Team:Penn/ProjectOverview'>Motivation</a></li><br />
<li><a href='/Team:Penn/ProjectResults'>Our Therapeutic</a></li><br />
<li><a href='/Team:Penn/Achievements'>Achievements</a></li><br />
<br />
<br />
<br />
</ul><br />
</li> <br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/LightActivatedOverview" style="font-size:13px;">Light Activation</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/LightActivatedOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/BLSensor'>YF1/FixJ BL Sensor</a></li><br />
<li><a href='/Team:Penn/LightActivatedLysis'>Light Activated Cell Lysis</a></li><br />
<br />
<br />
</ul><br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/SurfaceDisplayOverview" style="font-size:13px;">Surface Display</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/SurfaceDisplayOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/SurfaceDisplay'>INPNC Surface Display</a></li><br />
<li><a href='/Team:Penn/Targeting'>HER2 Targeting </a></li><br />
<li><a href='/Team:Penn/SurfaceDisplayBBa'>BBa Surface Display Platform</a></li><br />
<br />
</ul><br />
<br />
</li> <br />
<br />
<br />
<li class="dropdown"><br />
<br />
<a href="#" style="font-size:13px;">Wet Lab</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/Notebook'>Notebook</a></li><br />
<li><a href='/Team:Penn/Safety'>Safety</a></li><br />
<li><a href='/Team:Penn/Protocols'>Protocols</a></li><br />
<li><a href='/Team:Penn/Parts'>Parts</a></li><br />
<br />
</ul><br />
<br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/HumanPracticesOverview" style="font-size:13px;">Human Practices</a><br />
<br />
<ul><br />
<li><a href='/Team:Penn/HumanPracticesOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/Barriers'>Bacterial Therapeutics</a></li> <br />
<li><a href='/Team:Penn/Outreach'>Outreach</a></li><br />
<li><a href='/Team:Penn/Perceptions'>Public Perception</a></li><br />
<li><a href='/Team:Penn/Nissle'>E. Coli Nissle 1917</a></li><br />
<li><a href='/Team:Penn/VerifiGEM'>VerifiGEM</a></li><br />
</ul> <br />
<br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/Team" style="font-size:13px;">About Us</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/Team'>Team</a></li> <br />
<li><a href='/Team:Penn/Sponsors'>Sponsors</a></li><br />
</ul> <br />
</li><br />
<br />
<div class="arrow"></div><br />
<br />
</ul><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/Template/SiteTeam:Penn/Template/Site2012-10-27T04:04:29Z<p>Amurthur: </p>
<hr />
<div><html><br />
<head><br />
<!-- coded by Ashwin Amurthur --><br />
<!-- University of Pennsylvania iGEM team WIKI --><br />
<link rel="stylesheet" href='/Team:Penn/css/navigation?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/layerslider?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/sliderstyle?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/style2?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/columns?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/igemcss?action=raw&amp;ctype=text/css' /><br />
<script type="text/javascript" src='/Team:Penn/js/accordion?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/jquery-1.7.1.min.js?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/layersliderkreatura?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/cufon-replace.js?action=raw&amp;ctype=text/javascript'></script><br />
<br />
<title>Penn 2012 iGEM Wiki</title><br />
<br />
<style type="text/css"><br />
<br />
a.white:link {color:#FFF;} /* unvisited link */<br />
a.white:visited {color:#FFF;} /* visited link */<br />
a.white:hover {color:#FF;} /* mouse over link */<br />
a.white:active {color:#FFF;} /* selected link */ <br />
<br />
<br />
#top-section2 {<br />
width:1000px;<br />
border:0px;<br />
height:0px;<br />
padding-bottom:0px;<br />
background: #01256e;<br />
margin-bottom: 0px;<br />
margin-top: 0px;<br />
}<br />
<br />
<br />
</style><br />
<br />
</head><br />
<br />
<body><br />
<div id="top-section"><br />
<div id="p-logo2"><br />
<a href="https://2012.igem.org/Team:Penn"><img src='https://static.igem.org/mediawiki/2012/f/fa/Sponsor-Image-Wiki-Blue-Background_v2.gif' width="1000" height="175" usemap="#igemmap"></a><br />
<br />
<br />
<map name="igemmap" ><br />
<area shape="rect" coords="0,122,67,170" href="https://2012.igem.org" alt="" title="" /><br />
<area shape="rect" coords="913,131,995,170" href="http://www.upenn.edu" alt="" title="" /><br />
<area shape="rect" coords="998,173,1000,175" href="http://www.image-maps.com/index.php?aff=mapped_users_1201210022332192" alt="Image Map" title="Image Map" /><br />
</map><br />
<br />
<br />
</div><br />
<br />
</div><br />
</div><br />
<br />
<div style="margin-top:0px;margin-bottom:0px; padding-bottom:0px;"><br />
<a href="https://2012.igem.org/Team:Penn" style="padding:0px; margin:0px;"><br />
<img src="https://static.igem.org/mediawiki/2012/1/10/Bgblue.png" /></a><br />
</div><br />
<br />
<br />
<ul class="nav"><br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/ProjectOverview" style="font-size:13px;">Our Project</a><br />
<br />
<ul><br />
<li><a href='/Team:Penn/ProjectOverview'>Motivation</a></li><br />
<li><a href='/Team:Penn/ProjectResults'>Our Therapeutic</a></li><br />
<li><a href='/Team:Penn/Achievements'>Achievements</a></li><br />
<br />
<br />
<br />
</ul><br />
</li> <br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/LightActivatedOverview" style="font-size:13px;">Light Activation</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/LightActivatedOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/BLSensor'>YF1/FixJ BL Sensor</a></li><br />
<li><a href='/Team:Penn/LightActivatedLysis'>Light Activated Cell Lysis</a></li><br />
<br />
<br />
</ul><br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/SurfaceDisplayOverview" style="font-size:13px;">Surface Display</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/SurfaceDisplayOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/SurfaceDisplay'>INPNC Surface Display</a></li><br />
<li><a href='/Team:Penn/Targeting'>HER2 Targeting </a></li><br />
<li><a href='/Team:Penn/SurfaceDisplayBBa'>BBa Surface Display Platform</a></li><br />
<br />
</ul><br />
<br />
</li> <br />
<br />
<br />
<li class="dropdown"><br />
<br />
<a href="#" style="font-size:13px;">Wet Lab</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/Notebook'>Notebook</a></li><br />
<li><a href='/Team:Penn/Safety'>Safety</a></li><br />
<li><a href='/Team:Penn/Protocols'>Protocols</a></li><br />
<li><a href='/Team:Penn/Parts'>Parts</a></li><br />
<br />
</ul><br />
<br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/HPGeneral" style="font-size:13px;">Human Practices</a><br />
<br />
<ul><br />
<li><a href='/Team:Penn/HumanPracticesOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/Barriers'>Bacterial Therapeutics</a></li> <br />
<li><a href='/Team:Penn/Outreach'>Outreach</a></li><br />
<li><a href='/Team:Penn/Perceptions'>Public Perception</a></li><br />
<li><a href='/Team:Penn/Nissle'>E. Coli Nissle 1917</a></li><br />
<li><a href='/Team:Penn/VerifiGEM'>VerifiGEM</a></li><br />
</ul> <br />
<br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/Team" style="font-size:13px;">About Us</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/Team'>Team</a></li> <br />
<li><a href='/Team:Penn/Sponsors'>Sponsors</a></li><br />
</ul> <br />
</li><br />
<br />
<div class="arrow"></div><br />
<br />
</ul><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/Template/Site2Team:Penn/Template/Site22012-10-27T04:04:19Z<p>Amurthur: </p>
<hr />
<div><html><br />
<head><br />
<!-- coded by Ashwin Amurthur --><br />
<!-- University of Pennsylvania iGEM team WIKI --><br />
<link rel="stylesheet" href='/Team:Penn/css/navigation?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/layerslider?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/sliderstyle?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/style2?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/columns?action=raw&amp;ctype=text/css' /><br />
<link rel="stylesheet" href='/Team:Penn/css/igemcss?action=raw&amp;ctype=text/css' /><br />
<script type="text/javascript" src='/Team:Penn/js/accordion?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/jquery-1.7.1.min.js?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/layersliderkreatura?action=raw&amp;ctype=text/javascript'></script><br />
<script type="text/javascript" src='/Team:Penn/js/cufon-replace.js?action=raw&amp;ctype=text/javascript'></script><br />
<br />
<title>Penn 2012 iGEM Wiki</title><br />
<br />
<style type="text/css"><br />
<br />
a.white:link {color:#FFF;} /* unvisited link */<br />
a.white:visited {color:#FFF;} /* visited link */<br />
a.white:hover {color:#FF;} /* mouse over link */<br />
a.white:active {color:#FFF;} /* selected link */ <br />
<br />
<br />
#top-section2 {<br />
width:1000px;<br />
border:0px;<br />
height:0px;<br />
padding-bottom:0px;<br />
background: #01256e;<br />
margin-bottom: 0px;<br />
margin-top: 0px;<br />
}<br />
<br />
<br />
</style><br />
<br />
</head><br />
<br />
<body><br />
<div id="top-section"><br />
<div id="p-logo2"><br />
<a href="https://2012.igem.org/Team:Penn"><img src='https://static.igem.org/mediawiki/2012/f/fa/Sponsor-Image-Wiki-Blue-Background_v2.gif' width="1000" height="175" usemap="#igemmap"></a><br />
<br />
<br />
<map name="igemmap" ><br />
<area shape="rect" coords="0,122,67,170" href="https://2012.igem.org" alt="" title="" /><br />
<area shape="rect" coords="913,131,995,170" href="http://www.upenn.edu" alt="" title="" /><br />
<area shape="rect" coords="998,173,1000,175" href="http://www.image-maps.com/index.php?aff=mapped_users_1201210022332192" alt="Image Map" title="Image Map" /><br />
</map><br />
<br />
<br />
</div><br />
<br />
</div><br />
</div><br />
<br />
<div style="margin-top:0px;margin-bottom:0px; padding-bottom:0px;"><br />
<a href="https://2012.igem.org/Team:Penn" style="padding:0px; margin:0px;"><br />
<img src="https://static.igem.org/mediawiki/2012/1/10/Bgblue.png" /></a><br />
</div><br />
<br />
<br />
<ul class="nav"><br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/ProjectOverview" style="font-size:13px;">Our Project</a><br />
<br />
<ul><br />
<li><a href='/Team:Penn/ProjectOverview'>Motivation</a></li><br />
<li><a href='/Team:Penn/ProjectResults'>Our Therapeutic</a></li><br />
<li><a href='/Team:Penn/Achievements'>Achievements</a></li><br />
<br />
<br />
<br />
</ul><br />
</li> <br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/LightActivatedOverview" style="font-size:13px;">Light Activation</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/LightActivatedOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/BLSensor'>YF1/FixJ BL Sensor</a></li><br />
<li><a href='/Team:Penn/LightActivatedLysis'>Light Activated Cell Lysis</a></li><br />
<br />
<br />
</ul><br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/SurfaceDisplayOverview" style="font-size:13px;">Surface Display</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/SurfaceDisplayOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/SurfaceDisplay'>INPNC Surface Display</a></li><br />
<li><a href='/Team:Penn/Targeting'>HER2 Targeting </a></li><br />
<li><a href='/Team:Penn/SurfaceDisplayBBa'>BBa Surface Display Platform</a></li><br />
<br />
</ul><br />
<br />
</li> <br />
<br />
<br />
<li class="dropdown"><br />
<br />
<a href="#" style="font-size:13px;">Wet Lab</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/Notebook'>Notebook</a></li><br />
<li><a href='/Team:Penn/Safety'>Safety</a></li><br />
<li><a href='/Team:Penn/Protocols'>Protocols</a></li><br />
<li><a href='/Team:Penn/Parts'>Parts</a></li><br />
<br />
</ul><br />
<br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/HPGeneral" style="font-size:13px;">Human Practices</a><br />
<br />
<ul><br />
<li><a href='/Team:Penn/HumanPracticesOverview'>Overview</a></li><br />
<li><a href='/Team:Penn/Barriers'>Bacterial Therapeutics</a></li> <br />
<li><a href='/Team:Penn/Outreach'>Outreach</a></li><br />
<li><a href='/Team:Penn/Perceptions'>Public Perception</a></li><br />
<li><a href='/Team:Penn/Nissle'>E. Coli Nissle 1917</a></li><br />
<li><a href='/Team:Penn/VerifiGEM'>VerifiGEM</a></li><br />
</ul> <br />
<br />
</li><br />
<br />
<li class="dropdown"><br />
<br />
<a href="/Team:Penn/Team" style="font-size:13px;">About Us</a><br />
<br />
<ul><br />
<br />
<li><a href='/Team:Penn/Team'>Team</a></li> <br />
<li><a href='/Team:Penn/Sponsors'>Sponsors</a></li><br />
</ul> <br />
</li><br />
<br />
<div class="arrow"></div><br />
<br />
</ul><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/NissleTeam:Penn/Nissle2012-10-27T04:01:53Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<style type="text/css"><br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Use of the Minimally Immunogenic E. Coli Strain Nissle 1917</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"> We have seen how the complex interplay between public opinion and science innovation can drastically affect the adoption and success of a new technology, such as our team's bacterial drug delivery system. As pointed out earlier, pervading public opinion towards a bacterial therapeutic system such as the one developed by our team this year would most likely be negative. In an effort to address some of these issues, we have set out to investigate ways our system could be made more palatable to the general public. </p><br />
<p style="color:black;text-indent:30px;"> One recent concept that we have identified as gaining general acceptance within the general public is the incorporation of "probiotic" organisms into a daily diet. Many foods, such as yogurts now advertise the presence of "probiotic" bacteria and there are "probiotic" supplements containing live bacterial cultures as well. One particular probiotic, E. Coli Nissle 1917 has attracted attention not only from the public, but also from the scientific community, where its potential beneficial properties have been investigated. The Nissle strain is notable for its lack of virulence factors and decreased immunogenecity [1]. These traits are what make Nissle a popular probiotic. Nissle has also been found to preferentially colonize tumors, proliferating wildly in the borders between live and necrotic tissue, a highly desirable trait for any potential cancer treatment [2]. Additional investigation has demonstrated that intravenously administered Nissle exhibits a similar behavior in breast cancer mouse models, and expression of recombinant azurin prevented cancer metastasis in mice [4]. However, the therapeutic potential of Nissle is not limited to cancer treatment. Nissle is also capable of enhancing wound healing through recombinant expression of human epidermal Growth Factor in the epithelial linings of the body, as well as reducing modulating responses to allergens [5,6].</p><br />
<p style="color:black;text-indent:30px;"> Based on these properties, we believe that demonstrating that our drug delivery system can be implemented in Nissle 1917 would be the first step to addressing the potential hesitance that the public may have to bacterial based therapies. Because the chassis for our system is a probiotic, we can avoid not only the technical difficulties of ensuring that the host for our system is inherently safe, but also proactively address (or at least minimize) the initial "knee-jerk" reactions that many members of the general public may have to the idea of a bacterial therapeutic. Furthermore, In order to fully realize the potential of Nissle, it is important to be able to easily and consistently manipulate and change its genetic information. Therefore, we have produced and characterized a process for generating chemically competent Nissle 1917 that can be produced in any standard microbiology lab, allowing future iGEM teams to unlock Nissle 1917's full potential.<br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Data</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
As shown below, chemically competent Nissle 1917 bacteria produced by the lab was capable of taking up the pDawn-ClyA light-based cytolysis module of our system. Furthermore, these bacteria can begin to express genes encoded on the plasmid, such as the kanR gene, which confers resistance to the Kanamycin found in the LB plates.</p><br />
</p><div align="center"><br><br />
<img src="https://static.igem.org/mediawiki/2012/9/90/20121004033558!IMG_3309.JPG" width="600" height="400"></div><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
As shown below, chemically competent Nissle 1917 bacteria produced by the lab was capable of taking up the pDawn-mCherry plasmid modeled on the pDawn-ClyA light-based cytolysis module of our system. Furthermore, these bacteria exhibit blue light-dependent expression of mCherry, a red fluorescent protein, as seen on the right of the photo.</p><br />
</p><div align="center"><br><br />
<img src="https://static.igem.org/mediawiki/2012/thumb/7/72/Nissle-1917-pDawn-mCherry-10-1-2012.jpg/400px-Nissle-1917-pDawn-mCherry-10-1-2012.jpg" width="240" height="360"></div><br />
</div><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Blood Agar Experiments with pDawn-ClyA transformed in Nissle 1917 </div></b><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/e/e9/Nissle-ClyA-DARK.jpg" height="466.7" width="700" /><br />
<b><div style="text-align:center">Nissle pDawn-ClyA Dark</div></b><br />
</div><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/8/8d/ClyA-Nissle-Light.jpg" height="466.7" width="700" /><br />
<b><div style="text-align:center">Nissle pDawn-ClyA Light</div></b><br />
</div><br />
</div><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Production of Chemically Competent Nissle 1917 Cells</div></b><br />
<ol><br />
<li>Inoculate one colony from LB plate into 2 ml LB liquid medium. Shake at 37 °C<br />
overnight.</li><br />
<li>Inoculate 1-ml overnight cell culture into 100 ml LB medium (in a 500 ml flask).</li><br />
<li>Shake vigorously at 37 °C to OD600 ~0.25-0.3.</li><br />
<li>Chill the culture on ice for 15 min. Also make sure the 0.1M CaCl2<br />
solution and 0.1M CaCl2 plus 15% glycerol are on ice.</li><br />
<li>Centrifuge the cells for 10 min at 5000 g at 4°C.</li><br />
<li>Discard the medium and resuspend the cell pellet in 30-40 ml cold 0.1M CaCl2. Keep the cells on ice for 30 min.</li><br />
<li>Centrifuge the cells as above.</li><br />
<li>Remove the supernatant, and resuspend the cell pellet in 6 ml 0.1 M CaCl2<br />
solution plus 15% glycerol.</li><br />
<li>Pipet 0.4-0.5 ml of the cell suspension into sterile 1.5 ml micro-centrifuge tubes. Flash freeze these tubes in liquid nitrogen and then transfer them to the -80 C freezer.<br />
<ul><br />
<li><br />
Note: Successful transformations have occured with 100uL of cells + 1ug of DNA, however the efficency of cells made through this process is lower than that of Subcloning Efficency DH5a from Invitrogen. After flash freezing, competency of cells prepared through this protocol increases over time with additional storage time in -80°C for approximately 3 days.<br />
</li><br />
</ul><br />
</ol><br />
</div><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">References:</div></b><br><br />
<br />
[1] Grozdanov, L., U. Zahringer, G. Blum-Oehler, L. Brade, A. Henne, Y. A. Knirel, U.Schombel, J. Schulze, U. Sonnenborn, G. Gottschalk, J. Hacker, E. T. Rietschel, and U. Dobrindt. "A Single Nucleotide Exchange in the Wzy Gene Is Responsible for the Semirough O6 Lipopolysaccharide Phenotype and Serum Sensitivity of Escherichia Coli Strain Nissle 1917." Journal of Bacteriology 184.21 (2002): 5912-925. Print.<br><br />
<br><br />
[2] Stritzker, J., S. Weibel, P. Hill, T. Oelschlaeger, W. Goebel, and A. Szalay.<br />
<br />
"Tumor-specific Colonization, Tissue Distribution, and Gene Induction by Probiotic Escherichia Coli Nissle 1917 in Live Mice." International Journal of Medical Microbiology 297.3 (2007): 151-62. 19 Apr. 2007. Web. 29 Sept. 2012.<br><br />
<br><br />
[3] Weise, Christin, Yan Zhu, Dennis Ernst, Anja A. Kühl, and Margitta Worm. "Oral<br />
<br />
Administration of Escherichia Coli Nissle 1917 Prevents Allergen-induced Dermatitis in Mice." Experimental Dermatology 20.10 (2011): 805-09. 11 July 2011. Web. 29 Sept. 2012. <br><br />
<br><br />
[4] Zhang, Y., L. Xia, X. Zhang, X. Ding, F. Yan, and F. Wu. "Escherichia Coli Nissle 1917 Targets and Restrains Mouse B16 Melanoma and 4T1 Breast Tumor through the Expression of Azurin Protein." Applied Environmental Microbiology (n.d.): n. pag. 24 Aug. 2012. Web. 29 Sept. 2012.</div><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/NissleTeam:Penn/Nissle2012-10-27T04:00:30Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<style type="text/css"><br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Use of the Minimally Immunogenic E. Coli Strain Nissle 1917</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"> We have seen how the complex interplay between public opinion and science innovation can drastically affect the adoption and success of a new technology, such as our team's bacterial drug delivery system. As pointed out earlier, pervading public opinion towards a bacterial therapeutic system such as the one developed by our team this year would most likely be negative. In an effort to address some of these issues, we have set out to investigate ways our system could be made more palatable to the general public. </p><br />
<p style="color:black;text-indent:30px;"> One recent concept that we have identified as gaining general acceptance within the general public is the incorporation of "probiotic" organisms into a daily diet. Many foods, such as yogurts now advertise the presence of "probiotic" bacteria and there are "probiotic" supplements containing live bacterial cultures as well. One particular probiotic, E. Coli Nissle 1917 has attracted attention not only from the public, but also from the scientific community, where its potential beneficial properties have been investigated. The Nissle strain is notable for its lack of virulence factors and decreased immunogenecity [1]. These traits are what make Nissle a popular probiotic. Nissle has also been found to preferentially colonize tumors, proliferating wildly in the borders between live and necrotic tissue, a highly desirable trait for any potential cancer treatment [2]. Additional investigation has demonstrated that intravenously administered Nissle exhibits a similar behavior in breast cancer mouse models, and expression of recombinant azurin prevented cancer metastasis in mice [4]. However, the therapeutic potential of Nissle is not limited to cancer treatment. Nissle is also capable of enhancing wound healing through recombinant expression of human epidermal Growth Factor in the epithelial linings of the body, as well as reducing modulating responses to allergens [5,6].</p><br />
<p style="color:black;text-indent:30px;"> Based on these properties, we believe that demonstrating that our drug delivery system can be implemented in Nissle 1917 would be the first step to addressing the potential hesitance that the public may have to bacterial based therapies. Because the chassis for our system is a probiotic, we can avoid not only the technical difficulties of ensuring that the host for our system is inherently safe, but also proactively address (or at least minimize) the initial "knee-jerk" reactions that many members of the general public may have to the idea of a bacterial therapeutic. Furthermore, In order to fully realize the potential of Nissle, it is important to be able to easily and consistently manipulate and change its genetic information. Therefore, we have produced and characterized a process for generating chemically competent Nissle 1917 that can be produced in any standard microbiology lab, allowing future iGEM teams to unlock Nissle 1917's full potential.<br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Data</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
As shown below, chemically competent Nissle 1917 bacteria produced by the lab was capable of taking up the pDawn-ClyA light-based cytolysis module of our system. Furthermore, these bacteria can begin to express genes encoded on the plasmid, such as the kanR gene, which confers resistance to the Kanamycin found in the LB plates.</p><br />
</p><div align="center"><br><br />
<img src="https://static.igem.org/mediawiki/2012/9/90/20121004033558!IMG_3309.JPG" width="600" height="400"></div><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
As shown below, chemically competent Nissle 1917 bacteria produced by the lab was capable of taking up the pDawn-mCherry plasmid modeled on the pDawn-ClyA light-based cytolysis module of our system. Furthermore, these bacteria exhibit blue light-dependent expression of mCherry, a red fluorescent protein, as seen on the right of the photo.</p><br />
</p><div align="center"><br><br />
<img src="https://static.igem.org/mediawiki/2012/thumb/7/72/Nissle-1917-pDawn-mCherry-10-1-2012.jpg/400px-Nissle-1917-pDawn-mCherry-10-1-2012.jpg" width="240" height="360"></div><br />
</div><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Blood Agar Experiments with pDawn-ClyA transformed in Nissle 1917 </div></b><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/e/e9/Nissle-ClyA-DARK.jpg" height="466.7" width="700" /><br />
<b><div style="text-align:center">Dark</div></b><br />
</div><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/8/8d/ClyA-Nissle-Light.jpg" height="466.7" width="700" /><br />
<b><div style="text-align:center">Light</div></b><br />
</div><br />
</div><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Production of Chemically Competent Nissle 1917 Cells</div></b><br />
<ol><br />
<li>Inoculate one colony from LB plate into 2 ml LB liquid medium. Shake at 37 °C<br />
overnight.</li><br />
<li>Inoculate 1-ml overnight cell culture into 100 ml LB medium (in a 500 ml flask).</li><br />
<li>Shake vigorously at 37 °C to OD600 ~0.25-0.3.</li><br />
<li>Chill the culture on ice for 15 min. Also make sure the 0.1M CaCl2<br />
solution and 0.1M CaCl2 plus 15% glycerol are on ice.</li><br />
<li>Centrifuge the cells for 10 min at 5000 g at 4°C.</li><br />
<li>Discard the medium and resuspend the cell pellet in 30-40 ml cold 0.1M CaCl2. Keep the cells on ice for 30 min.</li><br />
<li>Centrifuge the cells as above.</li><br />
<li>Remove the supernatant, and resuspend the cell pellet in 6 ml 0.1 M CaCl2<br />
solution plus 15% glycerol.</li><br />
<li>Pipet 0.4-0.5 ml of the cell suspension into sterile 1.5 ml micro-centrifuge tubes. Flash freeze these tubes in liquid nitrogen and then transfer them to the -80 C freezer.<br />
<ul><br />
<li><br />
Note: Successful transformations have occured with 100uL of cells + 1ug of DNA, however the efficency of cells made through this process is lower than that of Subcloning Efficency DH5a from Invitrogen. After flash freezing, competency of cells prepared through this protocol increases over time with additional storage time in -80°C for approximately 3 days.<br />
</li><br />
</ul><br />
</ol><br />
</div><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">References:</div></b><br><br />
<br />
[1] Grozdanov, L., U. Zahringer, G. Blum-Oehler, L. Brade, A. Henne, Y. A. Knirel, U.Schombel, J. Schulze, U. Sonnenborn, G. Gottschalk, J. Hacker, E. T. Rietschel, and U. Dobrindt. "A Single Nucleotide Exchange in the Wzy Gene Is Responsible for the Semirough O6 Lipopolysaccharide Phenotype and Serum Sensitivity of Escherichia Coli Strain Nissle 1917." Journal of Bacteriology 184.21 (2002): 5912-925. Print.<br><br />
<br><br />
[2] Stritzker, J., S. Weibel, P. Hill, T. Oelschlaeger, W. Goebel, and A. Szalay.<br />
<br />
"Tumor-specific Colonization, Tissue Distribution, and Gene Induction by Probiotic Escherichia Coli Nissle 1917 in Live Mice." International Journal of Medical Microbiology 297.3 (2007): 151-62. 19 Apr. 2007. Web. 29 Sept. 2012.<br><br />
<br><br />
[3] Weise, Christin, Yan Zhu, Dennis Ernst, Anja A. Kühl, and Margitta Worm. "Oral<br />
<br />
Administration of Escherichia Coli Nissle 1917 Prevents Allergen-induced Dermatitis in Mice." Experimental Dermatology 20.10 (2011): 805-09. 11 July 2011. Web. 29 Sept. 2012. <br><br />
<br><br />
[4] Zhang, Y., L. Xia, X. Zhang, X. Ding, F. Yan, and F. Wu. "Escherichia Coli Nissle 1917 Targets and Restrains Mouse B16 Melanoma and 4T1 Breast Tumor through the Expression of Azurin Protein." Applied Environmental Microbiology (n.d.): n. pag. 24 Aug. 2012. Web. 29 Sept. 2012.</div><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/NissleTeam:Penn/Nissle2012-10-27T03:59:38Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<style type="text/css"><br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Use of the Minimally Immunogenic E. Coli Strain Nissle 1917</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"> We have seen how the complex interplay between public opinion and science innovation can drastically affect the adoption and success of a new technology, such as our team's bacterial drug delivery system. As pointed out earlier, pervading public opinion towards a bacterial therapeutic system such as the one developed by our team this year would most likely be negative. In an effort to address some of these issues, we have set out to investigate ways our system could be made more palatable to the general public. </p><br />
<p style="color:black;text-indent:30px;"> One recent concept that we have identified as gaining general acceptance within the general public is the incorporation of "probiotic" organisms into a daily diet. Many foods, such as yogurts now advertise the presence of "probiotic" bacteria and there are "probiotic" supplements containing live bacterial cultures as well. One particular probiotic, E. Coli Nissle 1917 has attracted attention not only from the public, but also from the scientific community, where its potential beneficial properties have been investigated. The Nissle strain is notable for its lack of virulence factors and decreased immunogenecity [1]. These traits are what make Nissle a popular probiotic. Nissle has also been found to preferentially colonize tumors, proliferating wildly in the borders between live and necrotic tissue, a highly desirable trait for any potential cancer treatment [2]. Additional investigation has demonstrated that intravenously administered Nissle exhibits a similar behavior in breast cancer mouse models, and expression of recombinant azurin prevented cancer metastasis in mice [4]. However, the therapeutic potential of Nissle is not limited to cancer treatment. Nissle is also capable of enhancing wound healing through recombinant expression of human epidermal Growth Factor in the epithelial linings of the body, as well as reducing modulating responses to allergens [5,6].</p><br />
<p style="color:black;text-indent:30px;"> Based on these properties, we believe that demonstrating that our drug delivery system can be implemented in Nissle 1917 would be the first step to addressing the potential hesitance that the public may have to bacterial based therapies. Because the chassis for our system is a probiotic, we can avoid not only the technical difficulties of ensuring that the host for our system is inherently safe, but also proactively address (or at least minimize) the initial "knee-jerk" reactions that many members of the general public may have to the idea of a bacterial therapeutic. Furthermore, In order to fully realize the potential of Nissle, it is important to be able to easily and consistently manipulate and change its genetic information. Therefore, we have produced and characterized a process for generating chemically competent Nissle 1917 that can be produced in any standard microbiology lab, allowing future iGEM teams to unlock Nissle 1917's full potential.<br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Data</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
As shown below, chemically competent Nissle 1917 bacteria produced by the lab was capable of taking up the pDawn-ClyA light-based cytolysis module of our system. Furthermore, these bacteria can begin to express genes encoded on the plasmid, such as the kanR gene, which confers resistance to the Kanamycin found in the LB plates.</p><br />
</p><div align="center"><br><br />
<img src="https://static.igem.org/mediawiki/2012/9/90/20121004033558!IMG_3309.JPG" width="600" height="400"></div><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
As shown below, chemically competent Nissle 1917 bacteria produced by the lab was capable of taking up the pDawn-mCherry plasmid modeled on the pDawn-ClyA light-based cytolysis module of our system. Furthermore, these bacteria exhibit blue light-dependent expression of mCherry, a red fluorescent protein, as seen on the right of the photo.</p><br />
</p><div align="center"><br><br />
<img src="https://static.igem.org/mediawiki/2012/thumb/7/72/Nissle-1917-pDawn-mCherry-10-1-2012.jpg/400px-Nissle-1917-pDawn-mCherry-10-1-2012.jpg" width="240" height="360"></div><br />
</div><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Blood Agar Experiments with pDawn-ClyA transformed in Nissle 1917 </div></b><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/e/e9/Nissle-ClyA-DARK.jpg" height="466.7" width="700" /><br />
</div><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/8/8d/ClyA-Nissle-Light.jpg" height="466.7" width="700" /><br />
</div><br />
</div><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Production of Chemically Competent Nissle 1917 Cells</div></b><br />
<ol><br />
<li>Inoculate one colony from LB plate into 2 ml LB liquid medium. Shake at 37 °C<br />
overnight.</li><br />
<li>Inoculate 1-ml overnight cell culture into 100 ml LB medium (in a 500 ml flask).</li><br />
<li>Shake vigorously at 37 °C to OD600 ~0.25-0.3.</li><br />
<li>Chill the culture on ice for 15 min. Also make sure the 0.1M CaCl2<br />
solution and 0.1M CaCl2 plus 15% glycerol are on ice.</li><br />
<li>Centrifuge the cells for 10 min at 5000 g at 4°C.</li><br />
<li>Discard the medium and resuspend the cell pellet in 30-40 ml cold 0.1M CaCl2. Keep the cells on ice for 30 min.</li><br />
<li>Centrifuge the cells as above.</li><br />
<li>Remove the supernatant, and resuspend the cell pellet in 6 ml 0.1 M CaCl2<br />
solution plus 15% glycerol.</li><br />
<li>Pipet 0.4-0.5 ml of the cell suspension into sterile 1.5 ml micro-centrifuge tubes. Flash freeze these tubes in liquid nitrogen and then transfer them to the -80 C freezer.<br />
<ul><br />
<li><br />
Note: Successful transformations have occured with 100uL of cells + 1ug of DNA, however the efficency of cells made through this process is lower than that of Subcloning Efficency DH5a from Invitrogen. After flash freezing, competency of cells prepared through this protocol increases over time with additional storage time in -80°C for approximately 3 days.<br />
</li><br />
</ul><br />
</ol><br />
</div><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">References:</div></b><br><br />
<br />
[1] Grozdanov, L., U. Zahringer, G. Blum-Oehler, L. Brade, A. Henne, Y. A. Knirel, U.Schombel, J. Schulze, U. Sonnenborn, G. Gottschalk, J. Hacker, E. T. Rietschel, and U. Dobrindt. "A Single Nucleotide Exchange in the Wzy Gene Is Responsible for the Semirough O6 Lipopolysaccharide Phenotype and Serum Sensitivity of Escherichia Coli Strain Nissle 1917." Journal of Bacteriology 184.21 (2002): 5912-925. Print.<br><br />
<br><br />
[2] Stritzker, J., S. Weibel, P. Hill, T. Oelschlaeger, W. Goebel, and A. Szalay.<br />
<br />
"Tumor-specific Colonization, Tissue Distribution, and Gene Induction by Probiotic Escherichia Coli Nissle 1917 in Live Mice." International Journal of Medical Microbiology 297.3 (2007): 151-62. 19 Apr. 2007. Web. 29 Sept. 2012.<br><br />
<br><br />
[3] Weise, Christin, Yan Zhu, Dennis Ernst, Anja A. Kühl, and Margitta Worm. "Oral<br />
<br />
Administration of Escherichia Coli Nissle 1917 Prevents Allergen-induced Dermatitis in Mice." Experimental Dermatology 20.10 (2011): 805-09. 11 July 2011. Web. 29 Sept. 2012. <br><br />
<br><br />
[4] Zhang, Y., L. Xia, X. Zhang, X. Ding, F. Yan, and F. Wu. "Escherichia Coli Nissle 1917 Targets and Restrains Mouse B16 Melanoma and 4T1 Breast Tumor through the Expression of Azurin Protein." Applied Environmental Microbiology (n.d.): n. pag. 24 Aug. 2012. Web. 29 Sept. 2012.</div><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/NissleTeam:Penn/Nissle2012-10-27T03:59:19Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<style type="text/css"><br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Use of the Minimally Immunogenic E. Coli Strain Nissle 1917</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"> We have seen how the complex interplay between public opinion and science innovation can drastically affect the adoption and success of a new technology, such as our team's bacterial drug delivery system. As pointed out earlier, pervading public opinion towards a bacterial therapeutic system such as the one developed by our team this year would most likely be negative. In an effort to address some of these issues, we have set out to investigate ways our system could be made more palatable to the general public. </p><br />
<p style="color:black;text-indent:30px;"> One recent concept that we have identified as gaining general acceptance within the general public is the incorporation of "probiotic" organisms into a daily diet. Many foods, such as yogurts now advertise the presence of "probiotic" bacteria and there are "probiotic" supplements containing live bacterial cultures as well. One particular probiotic, E. Coli Nissle 1917 has attracted attention not only from the public, but also from the scientific community, where its potential beneficial properties have been investigated. The Nissle strain is notable for its lack of virulence factors and decreased immunogenecity [1]. These traits are what make Nissle a popular probiotic. Nissle has also been found to preferentially colonize tumors, proliferating wildly in the borders between live and necrotic tissue, a highly desirable trait for any potential cancer treatment [2]. Additional investigation has demonstrated that intravenously administered Nissle exhibits a similar behavior in breast cancer mouse models, and expression of recombinant azurin prevented cancer metastasis in mice [4]. However, the therapeutic potential of Nissle is not limited to cancer treatment. Nissle is also capable of enhancing wound healing through recombinant expression of human epidermal Growth Factor in the epithelial linings of the body, as well as reducing modulating responses to allergens [5,6].</p><br />
<p style="color:black;text-indent:30px;"> Based on these properties, we believe that demonstrating that our drug delivery system can be implemented in Nissle 1917 would be the first step to addressing the potential hesitance that the public may have to bacterial based therapies. Because the chassis for our system is a probiotic, we can avoid not only the technical difficulties of ensuring that the host for our system is inherently safe, but also proactively address (or at least minimize) the initial "knee-jerk" reactions that many members of the general public may have to the idea of a bacterial therapeutic. Furthermore, In order to fully realize the potential of Nissle, it is important to be able to easily and consistently manipulate and change its genetic information. Therefore, we have produced and characterized a process for generating chemically competent Nissle 1917 that can be produced in any standard microbiology lab, allowing future iGEM teams to unlock Nissle 1917's full potential.<br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Data</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
As shown below, chemically competent Nissle 1917 bacteria produced by the lab was capable of taking up the pDawn-ClyA light-based cytolysis module of our system. Furthermore, these bacteria can begin to express genes encoded on the plasmid, such as the kanR gene, which confers resistance to the Kanamycin found in the LB plates.</p><br />
</p><div align="center"><br><br />
<img src="https://static.igem.org/mediawiki/2012/9/90/20121004033558!IMG_3309.JPG" width="600" height="400"></div><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
As shown below, chemically competent Nissle 1917 bacteria produced by the lab was capable of taking up the pDawn-mCherry plasmid modeled on the pDawn-ClyA light-based cytolysis module of our system. Furthermore, these bacteria exhibit blue light-dependent expression of mCherry, a red fluorescent protein, as seen on the right of the photo.</p><br />
</p><div align="center"><br><br />
<img src="https://static.igem.org/mediawiki/2012/thumb/7/72/Nissle-1917-pDawn-mCherry-10-1-2012.jpg/400px-Nissle-1917-pDawn-mCherry-10-1-2012.jpg" width="240" height="360"></div><br />
</div><br />
<br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Production of Chemically Competent Nissle 1917 Cells</div></b><br />
<ol><br />
<li>Inoculate one colony from LB plate into 2 ml LB liquid medium. Shake at 37 °C<br />
overnight.</li><br />
<li>Inoculate 1-ml overnight cell culture into 100 ml LB medium (in a 500 ml flask).</li><br />
<li>Shake vigorously at 37 °C to OD600 ~0.25-0.3.</li><br />
<li>Chill the culture on ice for 15 min. Also make sure the 0.1M CaCl2<br />
solution and 0.1M CaCl2 plus 15% glycerol are on ice.</li><br />
<li>Centrifuge the cells for 10 min at 5000 g at 4°C.</li><br />
<li>Discard the medium and resuspend the cell pellet in 30-40 ml cold 0.1M CaCl2. Keep the cells on ice for 30 min.</li><br />
<li>Centrifuge the cells as above.</li><br />
<li>Remove the supernatant, and resuspend the cell pellet in 6 ml 0.1 M CaCl2<br />
solution plus 15% glycerol.</li><br />
<li>Pipet 0.4-0.5 ml of the cell suspension into sterile 1.5 ml micro-centrifuge tubes. Flash freeze these tubes in liquid nitrogen and then transfer them to the -80 C freezer.<br />
<ul><br />
<li><br />
Note: Successful transformations have occured with 100uL of cells + 1ug of DNA, however the efficency of cells made through this process is lower than that of Subcloning Efficency DH5a from Invitrogen. After flash freezing, competency of cells prepared through this protocol increases over time with additional storage time in -80°C for approximately 3 days.<br />
</li><br />
</ul><br />
</ol><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Blood Agar Experiments with pDawn-ClyA transformed in Nissle 1917 </div></b><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/e/e9/Nissle-ClyA-DARK.jpg" height="466.7" width="700" /><br />
</div><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/8/8d/ClyA-Nissle-Light.jpg" height="466.7" width="700" /><br />
</div><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">References:</div></b><br><br />
<br />
[1] Grozdanov, L., U. Zahringer, G. Blum-Oehler, L. Brade, A. Henne, Y. A. Knirel, U.Schombel, J. Schulze, U. Sonnenborn, G. Gottschalk, J. Hacker, E. T. Rietschel, and U. Dobrindt. "A Single Nucleotide Exchange in the Wzy Gene Is Responsible for the Semirough O6 Lipopolysaccharide Phenotype and Serum Sensitivity of Escherichia Coli Strain Nissle 1917." Journal of Bacteriology 184.21 (2002): 5912-925. Print.<br><br />
<br><br />
[2] Stritzker, J., S. Weibel, P. Hill, T. Oelschlaeger, W. Goebel, and A. Szalay.<br />
<br />
"Tumor-specific Colonization, Tissue Distribution, and Gene Induction by Probiotic Escherichia Coli Nissle 1917 in Live Mice." International Journal of Medical Microbiology 297.3 (2007): 151-62. 19 Apr. 2007. Web. 29 Sept. 2012.<br><br />
<br><br />
[3] Weise, Christin, Yan Zhu, Dennis Ernst, Anja A. Kühl, and Margitta Worm. "Oral<br />
<br />
Administration of Escherichia Coli Nissle 1917 Prevents Allergen-induced Dermatitis in Mice." Experimental Dermatology 20.10 (2011): 805-09. 11 July 2011. Web. 29 Sept. 2012. <br><br />
<br><br />
[4] Zhang, Y., L. Xia, X. Zhang, X. Ding, F. Yan, and F. Wu. "Escherichia Coli Nissle 1917 Targets and Restrains Mouse B16 Melanoma and 4T1 Breast Tumor through the Expression of Azurin Protein." Applied Environmental Microbiology (n.d.): n. pag. 24 Aug. 2012. Web. 29 Sept. 2012.</div><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/NissleTeam:Penn/Nissle2012-10-27T03:59:06Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<style type="text/css"><br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Use of the Minimally Immunogenic E. Coli Strain Nissle 1917</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"> We have seen how the complex interplay between public opinion and science innovation can drastically affect the adoption and success of a new technology, such as our team's bacterial drug delivery system. As pointed out earlier, pervading public opinion towards a bacterial therapeutic system such as the one developed by our team this year would most likely be negative. In an effort to address some of these issues, we have set out to investigate ways our system could be made more palatable to the general public. </p><br />
<p style="color:black;text-indent:30px;"> One recent concept that we have identified as gaining general acceptance within the general public is the incorporation of "probiotic" organisms into a daily diet. Many foods, such as yogurts now advertise the presence of "probiotic" bacteria and there are "probiotic" supplements containing live bacterial cultures as well. One particular probiotic, E. Coli Nissle 1917 has attracted attention not only from the public, but also from the scientific community, where its potential beneficial properties have been investigated. The Nissle strain is notable for its lack of virulence factors and decreased immunogenecity [1]. These traits are what make Nissle a popular probiotic. Nissle has also been found to preferentially colonize tumors, proliferating wildly in the borders between live and necrotic tissue, a highly desirable trait for any potential cancer treatment [2]. Additional investigation has demonstrated that intravenously administered Nissle exhibits a similar behavior in breast cancer mouse models, and expression of recombinant azurin prevented cancer metastasis in mice [4]. However, the therapeutic potential of Nissle is not limited to cancer treatment. Nissle is also capable of enhancing wound healing through recombinant expression of human epidermal Growth Factor in the epithelial linings of the body, as well as reducing modulating responses to allergens [5,6].</p><br />
<p style="color:black;text-indent:30px;"> Based on these properties, we believe that demonstrating that our drug delivery system can be implemented in Nissle 1917 would be the first step to addressing the potential hesitance that the public may have to bacterial based therapies. Because the chassis for our system is a probiotic, we can avoid not only the technical difficulties of ensuring that the host for our system is inherently safe, but also proactively address (or at least minimize) the initial "knee-jerk" reactions that many members of the general public may have to the idea of a bacterial therapeutic. Furthermore, In order to fully realize the potential of Nissle, it is important to be able to easily and consistently manipulate and change its genetic information. Therefore, we have produced and characterized a process for generating chemically competent Nissle 1917 that can be produced in any standard microbiology lab, allowing future iGEM teams to unlock Nissle 1917's full potential.<br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Data</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
As shown below, chemically competent Nissle 1917 bacteria produced by the lab was capable of taking up the pDawn-ClyA light-based cytolysis module of our system. Furthermore, these bacteria can begin to express genes encoded on the plasmid, such as the kanR gene, which confers resistance to the Kanamycin found in the LB plates.</p><br />
</p><div align="center"><br><br />
<img src="https://static.igem.org/mediawiki/2012/9/90/20121004033558!IMG_3309.JPG" width="600" height="400"></div><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
As shown below, chemically competent Nissle 1917 bacteria produced by the lab was capable of taking up the pDawn-mCherry plasmid modeled on the pDawn-ClyA light-based cytolysis module of our system. Furthermore, these bacteria exhibit blue light-dependent expression of mCherry, a red fluorescent protein, as seen on the right of the photo.</p><br />
</p><div align="center"><br><br />
<img src="https://static.igem.org/mediawiki/2012/thumb/7/72/Nissle-1917-pDawn-mCherry-10-1-2012.jpg/400px-Nissle-1917-pDawn-mCherry-10-1-2012.jpg" width="240" height="360"></div><br />
</div><br />
<br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Production of Chemically Competent Nissle 1917 Cells</div></b><br />
<ol><br />
<li>Inoculate one colony from LB plate into 2 ml LB liquid medium. Shake at 37 °C<br />
overnight.</li><br />
<li>Inoculate 1-ml overnight cell culture into 100 ml LB medium (in a 500 ml flask).</li><br />
<li>Shake vigorously at 37 °C to OD600 ~0.25-0.3.</li><br />
<li>Chill the culture on ice for 15 min. Also make sure the 0.1M CaCl2<br />
solution and 0.1M CaCl2 plus 15% glycerol are on ice.</li><br />
<li>Centrifuge the cells for 10 min at 5000 g at 4°C.</li><br />
<li>Discard the medium and resuspend the cell pellet in 30-40 ml cold 0.1M CaCl2. Keep the cells on ice for 30 min.</li><br />
<li>Centrifuge the cells as above.</li><br />
<li>Remove the supernatant, and resuspend the cell pellet in 6 ml 0.1 M CaCl2<br />
solution plus 15% glycerol.</li><br />
<li>Pipet 0.4-0.5 ml of the cell suspension into sterile 1.5 ml micro-centrifuge tubes. Flash freeze these tubes in liquid nitrogen and then transfer them to the -80 C freezer.<br />
<ul><br />
<li><br />
Note: Successful transformations have occured with 100uL of cells + 1ug of DNA, however the efficency of cells made through this process is lower than that of Subcloning Efficency DH5a from Invitrogen. After flash freezing, competency of cells prepared through this protocol increases over time with additional storage time in -80°C for approximately 3 days.<br />
</li><br />
</ul><br />
</ol><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Blood Agar Experiments with pDawn-ClyA transformed in Nissle 1917 </div></b><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/e/e9/Nissle-ClyA-DARK.jpg" height="466.7" width="700" /><br />
</div><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/8/8d/ClyA-Nissle-Light.jpg" height="466.7" width="700" /><br />
</div><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">References:</div></b><br><br />
<br />
[1] Grozdanov, L., U. Zahringer, G. Blum-Oehler, L. Brade, A. Henne, Y. A. Knirel, U.Schombel, J. Schulze, U. Sonnenborn, G. Gottschalk, J. Hacker, E. T. Rietschel, and U. Dobrindt. "A Single Nucleotide Exchange in the Wzy Gene Is Responsible for the Semirough O6 Lipopolysaccharide Phenotype and Serum Sensitivity of Escherichia Coli Strain Nissle 1917." Journal of Bacteriology 184.21 (2002): 5912-925. Print.<br><br />
<br><br />
[2] Stritzker, J., S. Weibel, P. Hill, T. Oelschlaeger, W. Goebel, and A. Szalay.<br />
<br />
"Tumor-specific Colonization, Tissue Distribution, and Gene Induction by Probiotic Escherichia Coli Nissle 1917 in Live Mice." International Journal of Medical Microbiology 297.3 (2007): 151-62. 19 Apr. 2007. Web. 29 Sept. 2012.<br><br />
<br><br />
[3] Weise, Christin, Yan Zhu, Dennis Ernst, Anja A. Kühl, and Margitta Worm. "Oral<br />
<br />
Administration of Escherichia Coli Nissle 1917 Prevents Allergen-induced Dermatitis in Mice." Experimental Dermatology 20.10 (2011): 805-09. 11 July 2011. Web. 29 Sept. 2012. <br><br />
<br><br />
[4] Zhang, Y., L. Xia, X. Zhang, X. Ding, F. Yan, and F. Wu. "Escherichia Coli Nissle 1917 Targets and Restrains Mouse B16 Melanoma and 4T1 Breast Tumor through the Expression of Azurin Protein." Applied Environmental Microbiology (n.d.): n. pag. 24 Aug. 2012. Web. 29 Sept. 2012.</div><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/NissleTeam:Penn/Nissle2012-10-27T03:58:09Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<style type="text/css"><br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Use of the Minimally Immunogenic E. Coli Strain Nissle 1917</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"> We have seen how the complex interplay between public opinion and science innovation can drastically affect the adoption and success of a new technology, such as our team's bacterial drug delivery system. As pointed out earlier, pervading public opinion towards a bacterial therapeutic system such as the one developed by our team this year would most likely be negative. In an effort to address some of these issues, we have set out to investigate ways our system could be made more palatable to the general public. </p><br />
<p style="color:black;text-indent:30px;"> One recent concept that we have identified as gaining general acceptance within the general public is the incorporation of "probiotic" organisms into a daily diet. Many foods, such as yogurts now advertise the presence of "probiotic" bacteria and there are "probiotic" supplements containing live bacterial cultures as well. One particular probiotic, E. Coli Nissle 1917 has attracted attention not only from the public, but also from the scientific community, where its potential beneficial properties have been investigated. The Nissle strain is notable for its lack of virulence factors and decreased immunogenecity [1]. These traits are what make Nissle a popular probiotic. Nissle has also been found to preferentially colonize tumors, proliferating wildly in the borders between live and necrotic tissue, a highly desirable trait for any potential cancer treatment [2]. Additional investigation has demonstrated that intravenously administered Nissle exhibits a similar behavior in breast cancer mouse models, and expression of recombinant azurin prevented cancer metastasis in mice [4]. However, the therapeutic potential of Nissle is not limited to cancer treatment. Nissle is also capable of enhancing wound healing through recombinant expression of human epidermal Growth Factor in the epithelial linings of the body, as well as reducing modulating responses to allergens [5,6].</p><br />
<p style="color:black;text-indent:30px;"> Based on these properties, we believe that demonstrating that our drug delivery system can be implemented in Nissle 1917 would be the first step to addressing the potential hesitance that the public may have to bacterial based therapies. Because the chassis for our system is a probiotic, we can avoid not only the technical difficulties of ensuring that the host for our system is inherently safe, but also proactively address (or at least minimize) the initial "knee-jerk" reactions that many members of the general public may have to the idea of a bacterial therapeutic. Furthermore, In order to fully realize the potential of Nissle, it is important to be able to easily and consistently manipulate and change its genetic information. Therefore, we have produced and characterized a process for generating chemically competent Nissle 1917 that can be produced in any standard microbiology lab, allowing future iGEM teams to unlock Nissle 1917's full potential.<br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Data</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
As shown below, chemically competent Nissle 1917 bacteria produced by the lab was capable of taking up the pDawn-ClyA light-based cytolysis module of our system. Furthermore, these bacteria can begin to express genes encoded on the plasmid, such as the kanR gene, which confers resistance to the Kanamycin found in the LB plates.</p><br />
</p><div align="center"><br><br />
<img src="https://static.igem.org/mediawiki/2012/9/90/20121004033558!IMG_3309.JPG" width="600" height="400"></div><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
As shown below, chemically competent Nissle 1917 bacteria produced by the lab was capable of taking up the pDawn-mCherry plasmid modeled on the pDawn-ClyA light-based cytolysis module of our system. Furthermore, these bacteria exhibit blue light-dependent expression of mCherry, a red fluorescent protein, as seen on the right of the photo.</p><br />
</p><div align="center"><br><br />
<img src="https://static.igem.org/mediawiki/2012/thumb/7/72/Nissle-1917-pDawn-mCherry-10-1-2012.jpg/400px-Nissle-1917-pDawn-mCherry-10-1-2012.jpg" width="240" height="360"></div><br />
div class="name" align="center">Blood Agar Experiments with pDawn-ClyA transformed in Nissle 1917 </div></b><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/e/e9/Nissle-ClyA-DARK.jpg" height="466.7" width="700" /><br />
</div><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/8/8d/ClyA-Nissle-Light.jpg" height="466.7" width="700" /><br />
</div><br />
</div><br />
<br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Production of Chemically Competent Nissle 1917 Cells</div></b><br />
<ol><br />
<li>Inoculate one colony from LB plate into 2 ml LB liquid medium. Shake at 37 °C<br />
overnight.</li><br />
<li>Inoculate 1-ml overnight cell culture into 100 ml LB medium (in a 500 ml flask).</li><br />
<li>Shake vigorously at 37 °C to OD600 ~0.25-0.3.</li><br />
<li>Chill the culture on ice for 15 min. Also make sure the 0.1M CaCl2<br />
solution and 0.1M CaCl2 plus 15% glycerol are on ice.</li><br />
<li>Centrifuge the cells for 10 min at 5000 g at 4°C.</li><br />
<li>Discard the medium and resuspend the cell pellet in 30-40 ml cold 0.1M CaCl2. Keep the cells on ice for 30 min.</li><br />
<li>Centrifuge the cells as above.</li><br />
<li>Remove the supernatant, and resuspend the cell pellet in 6 ml 0.1 M CaCl2<br />
solution plus 15% glycerol.</li><br />
<li>Pipet 0.4-0.5 ml of the cell suspension into sterile 1.5 ml micro-centrifuge tubes. Flash freeze these tubes in liquid nitrogen and then transfer them to the -80 C freezer.<br />
<ul><br />
<li><br />
Note: Successful transformations have occured with 100uL of cells + 1ug of DNA, however the efficency of cells made through this process is lower than that of Subcloning Efficency DH5a from Invitrogen. After flash freezing, competency of cells prepared through this protocol increases over time with additional storage time in -80°C for approximately 3 days.<br />
</li><br />
</ul><br />
</ol><br />
<br />
<b><<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">References:</div></b><br><br />
<br />
[1] Grozdanov, L., U. Zahringer, G. Blum-Oehler, L. Brade, A. Henne, Y. A. Knirel, U.Schombel, J. Schulze, U. Sonnenborn, G. Gottschalk, J. Hacker, E. T. Rietschel, and U. Dobrindt. "A Single Nucleotide Exchange in the Wzy Gene Is Responsible for the Semirough O6 Lipopolysaccharide Phenotype and Serum Sensitivity of Escherichia Coli Strain Nissle 1917." Journal of Bacteriology 184.21 (2002): 5912-925. Print.<br><br />
<br><br />
[2] Stritzker, J., S. Weibel, P. Hill, T. Oelschlaeger, W. Goebel, and A. Szalay.<br />
<br />
"Tumor-specific Colonization, Tissue Distribution, and Gene Induction by Probiotic Escherichia Coli Nissle 1917 in Live Mice." International Journal of Medical Microbiology 297.3 (2007): 151-62. 19 Apr. 2007. Web. 29 Sept. 2012.<br><br />
<br><br />
[3] Weise, Christin, Yan Zhu, Dennis Ernst, Anja A. Kühl, and Margitta Worm. "Oral<br />
<br />
Administration of Escherichia Coli Nissle 1917 Prevents Allergen-induced Dermatitis in Mice." Experimental Dermatology 20.10 (2011): 805-09. 11 July 2011. Web. 29 Sept. 2012. <br><br />
<br><br />
[4] Zhang, Y., L. Xia, X. Zhang, X. Ding, F. Yan, and F. Wu. "Escherichia Coli Nissle 1917 Targets and Restrains Mouse B16 Melanoma and 4T1 Breast Tumor through the Expression of Azurin Protein." Applied Environmental Microbiology (n.d.): n. pag. 24 Aug. 2012. Web. 29 Sept. 2012.</div><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/NissleTeam:Penn/Nissle2012-10-27T03:57:37Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<style type="text/css"><br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Use of the Minimally Immunogenic E. Coli Strain Nissle 1917</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"> We have seen how the complex interplay between public opinion and science innovation can drastically affect the adoption and success of a new technology, such as our team's bacterial drug delivery system. As pointed out earlier, pervading public opinion towards a bacterial therapeutic system such as the one developed by our team this year would most likely be negative. In an effort to address some of these issues, we have set out to investigate ways our system could be made more palatable to the general public. </p><br />
<p style="color:black;text-indent:30px;"> One recent concept that we have identified as gaining general acceptance within the general public is the incorporation of "probiotic" organisms into a daily diet. Many foods, such as yogurts now advertise the presence of "probiotic" bacteria and there are "probiotic" supplements containing live bacterial cultures as well. One particular probiotic, E. Coli Nissle 1917 has attracted attention not only from the public, but also from the scientific community, where its potential beneficial properties have been investigated. The Nissle strain is notable for its lack of virulence factors and decreased immunogenecity [1]. These traits are what make Nissle a popular probiotic. Nissle has also been found to preferentially colonize tumors, proliferating wildly in the borders between live and necrotic tissue, a highly desirable trait for any potential cancer treatment [2]. Additional investigation has demonstrated that intravenously administered Nissle exhibits a similar behavior in breast cancer mouse models, and expression of recombinant azurin prevented cancer metastasis in mice [4]. However, the therapeutic potential of Nissle is not limited to cancer treatment. Nissle is also capable of enhancing wound healing through recombinant expression of human epidermal Growth Factor in the epithelial linings of the body, as well as reducing modulating responses to allergens [5,6].</p><br />
<p style="color:black;text-indent:30px;"> Based on these properties, we believe that demonstrating that our drug delivery system can be implemented in Nissle 1917 would be the first step to addressing the potential hesitance that the public may have to bacterial based therapies. Because the chassis for our system is a probiotic, we can avoid not only the technical difficulties of ensuring that the host for our system is inherently safe, but also proactively address (or at least minimize) the initial "knee-jerk" reactions that many members of the general public may have to the idea of a bacterial therapeutic. Furthermore, In order to fully realize the potential of Nissle, it is important to be able to easily and consistently manipulate and change its genetic information. Therefore, we have produced and characterized a process for generating chemically competent Nissle 1917 that can be produced in any standard microbiology lab, allowing future iGEM teams to unlock Nissle 1917's full potential.<br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Data</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
As shown below, chemically competent Nissle 1917 bacteria produced by the lab was capable of taking up the pDawn-ClyA light-based cytolysis module of our system. Furthermore, these bacteria can begin to express genes encoded on the plasmid, such as the kanR gene, which confers resistance to the Kanamycin found in the LB plates.</p><br />
</p><div align="center"><br><br />
<img src="https://static.igem.org/mediawiki/2012/9/90/20121004033558!IMG_3309.JPG" width="600" height="400"></div><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
As shown below, chemically competent Nissle 1917 bacteria produced by the lab was capable of taking up the pDawn-mCherry plasmid modeled on the pDawn-ClyA light-based cytolysis module of our system. Furthermore, these bacteria exhibit blue light-dependent expression of mCherry, a red fluorescent protein, as seen on the right of the photo.</p><br />
</p><div align="center"><br><br />
<img src="https://static.igem.org/mediawiki/2012/thumb/7/72/Nissle-1917-pDawn-mCherry-10-1-2012.jpg/400px-Nissle-1917-pDawn-mCherry-10-1-2012.jpg" width="240" height="360"></div><br />
</div><br />
<br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Production of Chemically Competent Nissle 1917 Cells</div></b><br />
<ol><br />
<li>Inoculate one colony from LB plate into 2 ml LB liquid medium. Shake at 37 °C<br />
overnight.</li><br />
<li>Inoculate 1-ml overnight cell culture into 100 ml LB medium (in a 500 ml flask).</li><br />
<li>Shake vigorously at 37 °C to OD600 ~0.25-0.3.</li><br />
<li>Chill the culture on ice for 15 min. Also make sure the 0.1M CaCl2<br />
solution and 0.1M CaCl2 plus 15% glycerol are on ice.</li><br />
<li>Centrifuge the cells for 10 min at 5000 g at 4°C.</li><br />
<li>Discard the medium and resuspend the cell pellet in 30-40 ml cold 0.1M CaCl2. Keep the cells on ice for 30 min.</li><br />
<li>Centrifuge the cells as above.</li><br />
<li>Remove the supernatant, and resuspend the cell pellet in 6 ml 0.1 M CaCl2<br />
solution plus 15% glycerol.</li><br />
<li>Pipet 0.4-0.5 ml of the cell suspension into sterile 1.5 ml micro-centrifuge tubes. Flash freeze these tubes in liquid nitrogen and then transfer them to the -80 C freezer.<br />
<ul><br />
<li><br />
Note: Successful transformations have occured with 100uL of cells + 1ug of DNA, however the efficency of cells made through this process is lower than that of Subcloning Efficency DH5a from Invitrogen. After flash freezing, competency of cells prepared through this protocol increases over time with additional storage time in -80°C for approximately 3 days.<br />
</li><br />
</ul><br />
</ol><br />
<br />
<b><div class="name" align="center">Blood Agar Experiments with pDawn-ClyA transformed in Nissle 1917 </div></b><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/e/e9/Nissle-ClyA-DARK.jpg" height="466.7" width="700" /><br />
</div><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/8/8d/ClyA-Nissle-Light.jpg" height="466.7" width="700" /><br />
</div><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">References:</div></b><br><br />
<br />
[1] Grozdanov, L., U. Zahringer, G. Blum-Oehler, L. Brade, A. Henne, Y. A. Knirel, U.Schombel, J. Schulze, U. Sonnenborn, G. Gottschalk, J. Hacker, E. T. Rietschel, and U. Dobrindt. "A Single Nucleotide Exchange in the Wzy Gene Is Responsible for the Semirough O6 Lipopolysaccharide Phenotype and Serum Sensitivity of Escherichia Coli Strain Nissle 1917." Journal of Bacteriology 184.21 (2002): 5912-925. Print.<br><br />
<br><br />
[2] Stritzker, J., S. Weibel, P. Hill, T. Oelschlaeger, W. Goebel, and A. Szalay.<br />
<br />
"Tumor-specific Colonization, Tissue Distribution, and Gene Induction by Probiotic Escherichia Coli Nissle 1917 in Live Mice." International Journal of Medical Microbiology 297.3 (2007): 151-62. 19 Apr. 2007. Web. 29 Sept. 2012.<br><br />
<br><br />
[3] Weise, Christin, Yan Zhu, Dennis Ernst, Anja A. Kühl, and Margitta Worm. "Oral<br />
<br />
Administration of Escherichia Coli Nissle 1917 Prevents Allergen-induced Dermatitis in Mice." Experimental Dermatology 20.10 (2011): 805-09. 11 July 2011. Web. 29 Sept. 2012. <br><br />
<br><br />
[4] Zhang, Y., L. Xia, X. Zhang, X. Ding, F. Yan, and F. Wu. "Escherichia Coli Nissle 1917 Targets and Restrains Mouse B16 Melanoma and 4T1 Breast Tumor through the Expression of Azurin Protein." Applied Environmental Microbiology (n.d.): n. pag. 24 Aug. 2012. Web. 29 Sept. 2012.</div><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/VerifiGEMTeam:Penn/VerifiGEM2012-10-27T03:53:57Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<style type="text/css"><br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<div class="bigbox"><br />
<b><div class="name" align="center">The Question: How Can We Increase the Accuracy of Biobrick Parts Submitted to the Registry?</div></b><br />
<br><br><br />
<b><div class="name" align="center">Our Solution: VerifiGEM</div></b><br />
</div><br />
<div style="padding-left:150px;"><br />
<div class="prezi-player"><style type="text/css" media="screen">.prezi-player { width: 700px; } .prezi-player-links { text-align: center; }</style><object id="prezi_xlruzy2n7spo" name="prezi_xlruzy2n7spo" classid="clsid:D27CDB6E-AE6D-11cf-96B8-444553540000" width="700" height="500"><param name="movie" value="http://prezi.com/bin/preziloader.swf"/><param name="allowfullscreen" value="true"/><param name="allowFullScreenInteractive" value="true"/><param name="allowscriptaccess" value="always"/><param name="wmode" value="direct"/><param name="bgcolor" value="#ffffff"/><param name="flashvars" value="prezi_id=xlruzy2n7spo&amp;lock_to_path=0&amp;color=ffffff&amp;autoplay=no&amp;autohide_ctrls=0"/><embed id="preziEmbed_xlruzy2n7spo" name="preziEmbed_xlruzy2n7spo" src="http://prezi.com/bin/preziloader.swf" type="application/x-shockwave-flash" allowfullscreen="true" allowFullScreenInteractive="true" allowscriptaccess="always" width="700" height="500" bgcolor="#ffffff" flashvars="prezi_id=xlruzy2n7spo&amp;lock_to_path=0&amp;color=ffffff&amp;autoplay=no&amp;autohide_ctrls=0"></embed></object><div class="prezi-player-links"><p><a title="VerifiGEM " href="http://prezi.com/xlruzy2n7spo/verifigem/"></a><a href="http://prezi.com"></a></p></div></div></div><br />
<br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/BLSensorTeam:Penn/BLSensor2012-10-27T03:53:00Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<br />
<br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">YF1/FixJ (pDawn) Objectives </div></b><br />
<br><br />
To characterize our pDawn gene expression system, we showed the following:<br />
<ol><br />
<li> pDawn allows for light-dependent gene expression in bacteria<br />
<li> pDawn allows for light-dependent lysis of mammalian cells by bacteria<br />
</ol><br />
<br />
</div><br />
<br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Light Dependent Gene Expression in Bacteria</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">We tested for light dependent gene expression by cloning in an mCherry reporter protein into the multiple cloning site of the pDawn system. First, we tested for the on-off ratio by growing cultures of BL21-pDawn-mCherry in both inducing and non-inducing conditions for 22 hours. After spinning down the cultures in a centrifuge, we were able to visually confirm the expression of mCherry due to the bacterial pellet grown in inducing conditions to be colored red, while the other pellet had no color (Figure 1).</p><br />
<br />
<div align="center"><br />
<br />
<img src="https://static.igem.org/mediawiki/2012/f/f4/Clark_Park_4.JPG" width="180" height="300"><br />
<br />
</div><br />
<br><br />
<div style="text-align:center"><b>Figure 1</b><br /></div><br />
</div><br />
<br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Characterizing Time-Dependent Gene Expression</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
We then characterized the induction kinetics of the pDawn system through an mCherry expression time course. We induced cultures of BL21 pDawn-mCherry for 0 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, 8 hours, or 22 hours in a 37C incubator shaking at 225 rpm, and then transferred them into a dark incubator under the same condition for the remaining growth period. After 24 hours, mCherry fluorescence was read on a Tecan Infinite m200 plate reader and normalized by OD. The cultures were then spun down in a centrifuge. These results can be seen in Figure 2. </p><br />
<br><br />
<div align="center"><br />
<br />
<img src="https://static.igem.org/mediawiki/2012/2/2a/PDawn-mCherry-Timecourse.gif" width="500"><br />
<br />
<img src="https://static.igem.org/mediawiki/2012/b/b9/Timecourse.png" width="500" ><br />
<br />
<br />
</div><br />
<br><br />
<div style="text-align:center"><b>Figure 2</b><br /></div><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">pDawn and Nissle 1917</div></b><br /><br />
<br />
<p style="color:black;text-indent:30px;"><br />
In order to further develop our system for future in vivo therapeutic applications, we transformed Nissle 1917 with pDawn-mCherry to see if we could implement our system into a non-pathogenic strain of E. coli. We repeated our initial experiments and achieved light-dependent gene expression in Nissle 1917 for the first time ever. We have also been able to transform our pDawn-ClyA construct into bacteria for use in blood agar plate experiments. <a href="https://2012.igem.org/Team:Penn/Nissle">Check it out! </a><br />
</p><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/7/72/Nissle-1917-pDawn-mCherry-10-1-2012.jpg" width="250" height="350"><br><br />
<br><br />
<b>Figure 3</b></div></div><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/BLSensorTeam:Penn/BLSensor2012-10-27T03:52:27Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<br />
<br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">YF1/FixJ (pDawn) Objectives </div></b><br />
<br><br />
To characterize our pDawn gene expression system, we showed the following:<br />
<ol><br />
<li> pDawn allows for light-dependent gene expression in bacteria<br />
<li> pDawn allows for light-dependent lysis of mammalian cells by bacteria<br />
</ol><br />
<br />
</div><br />
<br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Light Dependent Gene Expression in Bacteria</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">We tested for light dependent gene expression by cloning in an mCherry reporter protein into the multiple cloning site of the pDawn system. First, we tested for the on-off ratio by growing cultures of BL21-pDawn-mCherry in both inducing and non-inducing conditions for 22 hours. After spinning down the cultures in a centrifuge, we were able to visually confirm the expression of mCherry due to the bacterial pellet grown in inducing conditions to be colored red, while the other pellet had no color (Figure 1).</p><br />
<br />
<div align="center"><br />
<br />
<img src="https://static.igem.org/mediawiki/2012/f/f4/Clark_Park_4.JPG" width="180" height="300"><br />
<br />
</div><br />
<br><br />
<div style="text-align:center"><b>Figure 1</b><br /></div><br />
</div><br />
<br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Characterizing Time-Dependent Gene Expression</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;"><br />
We then characterized the induction kinetics of the pDawn system through an mCherry expression time course. We induced cultures of BL21 pDawn-mCherry for 0 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, 8 hours, or 22 hours in a 37C incubator shaking at 225 rpm, and then transferred them into a dark incubator under the same condition for the remaining growth period. After 24 hours, mCherry fluorescence was read on a Tecan Infinite m200 plate reader and normalized by OD. The cultures were then spun down in a centrifuge. These results can be seen in Figure 2. </p><br />
<br><br />
<div align="center"><br />
<br />
<img src="https://static.igem.org/mediawiki/2012/2/2a/PDawn-mCherry-Timecourse.gif" width="500"><br />
<br />
<img src="https://static.igem.org/mediawiki/2012/b/b9/Timecourse.png" width="500" ><br />
<br />
<br />
</div><br />
<br><br />
<div style="text-align:center"><b>Figure 2</b><br /></div><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">pDawn and Nissle 1917</div></b><br /><br />
<br />
<p style="color:black;text-indent:30px;"><br />
In order to further develop our system for future in vivo therapeutic applications, we transformed Nissle 1917 with pDawn-mCherry to see if we could implement our system into a non-pathogenic strain of E. coli. We repeated our initial experiments and achieved light-dependent gene expression in Nissle 1917 for the first time ever. We have also been able to transform our pDawn-ClyA construct into bacteria for use in blood agar plate experiments. <a href="https://2012.igem.org/Team:Penn/Nissle">Check it out </a><br />
</p><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/7/72/Nissle-1917-pDawn-mCherry-10-1-2012.jpg" width="250" height="350"><br><br />
<br><br />
<b>Figure 3</b></div></div><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/ProjectResultsTeam:Penn/ProjectResults2012-10-27T03:48:54Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">A Novel Therapeutic Platform</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">What if you could combine spatial targeting and cellular targeting into the same therapeutic? This idea is unprecedented but would allow for precise targeting of specific cells within a specific area, leaving healthy tissue intact and keeping side effects to a minimum. Higher dose precision means more of the therapeutic would be used efficiently in the targeted area and the dependency on passive diffusion – and the uncertainties that comes with it – would be eliminated.</p><br />
<br />
<p style="color:black;text-indent:30px;">The 2012 Penn iGEM team has engineered a novel, <b>modular</b> platform for targeted therapeutics that employs simultaneous spatial and cellular targeting. We have achieved spatial (and temporal) targeting with a blue light-switchable transgene expression system and cellular targeting through display of an antibody-mimetic protein on the surface of E. coli for the first time. Our platform also enables more precise dose control in the targeted area through the length of blue-light exposure, which allows us to regulate effective levels of transgene expression.</p><br />
<br />
<p style="color:black;text-indent:30px;">As a proof of concept, we applied our system to the treatment of cancer, a disease in which spatial and cellular targeting are of utmost importance. We displayed a high-affinity antibody-mimetic protein that targets Human Epidermal Growth Factor Receptor 2 (HER2), a protein commonly overexpressed in cancer cells, especially in breast cancer tumors. We combined this cellular targeting with a light-activated cytotoxic protein delivery system to successfully target and kill breast cancer cells.</p><br />
</div><br />
<br />
<div style="background-color:#01256e;" align="center"><br />
<iframe width="700" height="393" style="margin-left:10px;padding:10px; background-color:#000000;"src="http://www.youtube.com/embed/0fqLD4IJMJo" frameborder="1" allowfullscreen></iframe><br />
</div> <br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Modularity</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">The strength in our platform lies in its modularity. Both the light-induced transgene expression system and the surface display system work and exist independently of each other on two compatible plasmids. These plasmids can be modified to meet the needs of any synthetic biologist. </p><br />
<br />
<p style="color:black;text-indent:30px;">Any gene of interest can be cloned into the light-induced transgene expression system and will then be expressed in a light-dependent and spatially controlled manner. Any targeting protein can be cloned into our surface display platform to allow cellular targeting against any desired biomarker. These modular plasmids may then be co-transformed together to create a bacterial therapeutic for a desired disease.</p><br />
<br />
<p style="color:black;text-indent:30px;">These modular components could also be extended into applications other than medical therapeutics, such as biocatalysis, manufacturing, and alternative energy. </p><br />
<br />
<br />
<b><div class="name" align="center">Generic Components</div></b><br />
<br />
<div align="center"><br />
<table width="860" cellspacing="20" style="background-color:#d7dce1;"><br />
<tr><br />
<td width="410"><img src="https://static.igem.org/mediawiki/2012/e/e9/Lightactivationmodule.gif" width="400" height="300" /><br />
</td><br />
<td width="410"><img src="https://static.igem.org/mediawiki/2012/5/5c/Surfdispmodule.gif"width = "400" height = "300" /><br />
</td><br />
</tr><br />
<tr valign="top"><br />
<td ><br />
<p style="text-align:justify;"><b>Spatial Targeting:</b> Surgeons excise a tumor manually, without regard for cellular heterogeneity within and around the tumor area.</p><br />
</td><br />
<td><br />
<p style="text-align:justify;"><b>Cellular Targeting:</b> Monoclonal antibodies identify antigens on certain cells or viruses. Monoclonal antibodies are often coupled with therapeutic agents. However, if the antigen is present in healthy tissue outside the diseased area, it will be targeted as well.</p><br />
</td><br />
</tr><br />
<br />
</table></div><br />
</div><br />
<br />
<br />
<br />
<br />
<br />
<div class="bigbox"><br />
<br />
<br />
<br />
<br />
<b><div class="name" align="center">Proof of Concept</div></b><br />
<br><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/f/f9/Full_Color_Complete_Schematic.gif" width="700" height="525" /></div><br><br />
<p style="color:black;text-indent:30px;"> For our engineered bacterial therapeutic, we chose to target breast cancer as a proof of concept using a blue light gene expression system. We chose this specific blue-light inducible gene expression system because of the well-characterized nature of the parts and the high on:off ratio.</p><br />
<br />
<p style="color:black;text-indent:30px;"> We also identified a well-characterized class of antibody mimetic proteins called designed ankryin repeat proteins, or DARPins. One DARPin in particular (H10-2-G3) was engineered to bind to the Human Epidermal Growth Factor 2 (HER2) at picomolar affinities. HER2 is overexpressed in breast cancer cells, and we had access to cell lines that overexpressed HER2 on their cell surface which we could use for binding assays. </p><br />
<br />
</div><br />
<br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Human Practices</div></b><br />
<br><br />
<br />
<p style="color:black;text-indent:30px;">Upon conception of this project, we realized that although hundreds of academic research projects and iGEM projects have been conducted in the realm of Health and Medicine, almost no engineered bacterial therapeutics have been brought to the clinic. We analyzed the hurdles and road ahead for bacterial synthetic biology-enabled therapeutics, compiling a thorough report with specific actions that iGEM teams in Health/Medicine can take to make their therapies more clinically tractable. This project directly informed our wet lab work, leading us to port our therapeutic system into a non-pathogenic, probiotic bacterial strain which is already used in human therapies today.</p><br />
<br />
<br />
<p style="color:black;text-indent:30px;">We hope our targeted therapeutic platform will allow other scientists and iGEM teams to target any cells they choose. In the near term, we are planning to test our cancer cell targeting/killing bacterial system in a mouse model and make a real impact on cancer research and therapy.</p><br />
</div></div>Amurthurhttp://2012.igem.org/Team:Penn/ProjectResultsTeam:Penn/ProjectResults2012-10-27T03:48:15Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">A Novel Therapeutic Platform</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">What if you could combine spatial targeting and cellular targeting into the same therapeutic? This idea is unprecedented but would allow for precise targeting of specific cells within a specific area, leaving healthy tissue intact and keeping side effects to a minimum. Higher dose precision means more of the therapeutic would be used efficiently in the targeted area and the dependency on passive diffusion – and the uncertainties that comes with it – would be eliminated.</p><br />
<br />
<p style="color:black;text-indent:30px;">The 2012 Penn iGEM team has engineered a novel, <b>modular</b> platform for targeted therapeutics that employs simultaneous spatial and cellular targeting. We have achieved spatial (and temporal) targeting with a blue light-switchable transgene expression system and cellular targeting through display of an antibody-mimetic protein on the surface of E. coli for the first time. Our platform also enables more precise dose control in the targeted area through the length of blue-light exposure, which allows us to regulate effective levels of transgene expression.</p><br />
<br />
<p style="color:black;text-indent:30px;">As a proof of concept, we applied our system to the treatment of cancer, a disease in which spatial and cellular targeting are of utmost importance. We displayed a high-affinity antibody-mimetic protein that targets Human Epidermal Growth Factor Receptor 2 (HER2), a protein commonly overexpressed in cancer cells, especially in breast cancer tumors. We combined this cellular targeting with a light-activated cytotoxic protein delivery system to successfully target and kill breast cancer cells.</p><br />
</div><br />
<br />
<div style="background-color:#01256e;" align="center"><br />
<iframe width="700" height="393" style="margin-left:10px;padding:10px; background-color:#000000;"src="http://www.youtube.com/embed/0fqLD4IJMJo" frameborder="1" allowfullscreen></iframe><br />
</div> <br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Modularity</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">The strength in our platform lies in its modularity. Both the light-induced transgene expression system and the surface display system work and exist independently of each other on two compatible plasmids. These plasmids can be modified to meet the needs of any synthetic biologist. </p><br />
<br />
<p style="color:black;text-indent:30px;">Any gene of interest can be cloned into the light-induced transgene expression system and will then be expressed in a light-dependent and spatially controlled manner. Any targeting protein can be cloned into our surface display platform to allow cellular targeting against any desired biomarker. These modular plasmids may then be co-transformed together to create a bacterial therapeutic for a desired disease.</p><br />
<br />
<p style="color:black;text-indent:30px;">These modular components could also be extended into applications other than medical therapeutics, such as biocatalysis, manufacturing, and alternative energy. </p><br />
<br />
<br />
<b><div class="name" align="center">Generic Components</div></b><br />
<br />
<div align="center"><br />
<table width="860" cellspacing="20" style="background-color:#d7dce1;"><br />
<tr><br />
<td width="410"><img src="https://static.igem.org/mediawiki/2012/e/e9/Lightactivationmodule.gif" width="400" height="300" /><br />
</td><br />
<td width="410"><img src="https://static.igem.org/mediawiki/2012/5/5c/Surfdispmodule.gif"width = "400" height = "300" /><br />
</td><br />
</tr><br />
<tr valign="top"><br />
<td ><br />
<p style="text-align:justify;"><b>Spatial Targeting:</b> Surgeons excise a tumor manually, without regard for cellular heterogeneity within and around the tumor area.</p><br />
</td><br />
<td><br />
<p style="text-align:justify;"><b>Cellular Targeting:</b> Monoclonal antibodies identify antigens on certain cells or viruses. Monoclonal antibodies are often coupled with therapeutic agents. However, if the antigen is present in healthy tissue outside the diseased area, it will be targeted as well.</p><br />
</td><br />
</tr><br />
<br />
</table></div><br />
</div><br />
<br />
<br />
<br />
<br />
<br />
<div class="bigbox"><br />
<br />
<br />
<br />
<br />
<b><div class="name" align="center">Proof of Concept</div></b><br />
<br><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/f/f9/Full_Color_Complete_Schematic.gif" width="700" height="525" /></div><br />
<p style="color:black;text-indent:30px;"> For our engineered bacterial therapeutic, we chose to target breast cancer as a proof of concept using a blue light gene expression system. We chose this specific blue-light inducible gene expression system because of the well-characterized nature of the parts and the high on:off ratio.</p><br />
<br />
<p style="color:black;text-indent:30px;"> We also identified a well-characterized class of antibody mimetic proteins called designed ankryin repeat proteins, or DARPins. One DARPin in particular (H10-2-G3) was engineered to bind to the Human Epidermal Growth Factor 2 (HER2) at picomolar affinities. HER2 is overexpressed in breast cancer cells, and we had access to cell lines that overexpressed HER2 on their cell surface which we could use for binding assays. </p><br />
<br />
</div><br />
<br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Human Practices</div></b><br />
<br><br />
<br />
<p style="color:black;text-indent:30px;">Upon conception of this project, we realized that although hundreds of academic research projects and iGEM projects have been conducted in the realm of Health and Medicine, almost no engineered bacterial therapeutics have been brought to the clinic. We analyzed the hurdles and road ahead for bacterial synthetic biology-enabled therapeutics, compiling a thorough report with specific actions that iGEM teams in Health/Medicine can take to make their therapies more clinically tractable. This project directly informed our wet lab work, leading us to port our therapeutic system into a non-pathogenic, probiotic bacterial strain which is already used in human therapies today.</p><br />
<br />
<br />
<p style="color:black;text-indent:30px;">We hope our targeted therapeutic platform will allow other scientists and iGEM teams to target any cells they choose. In the near term, we are planning to test our cancer cell targeting/killing bacterial system in a mouse model and make a real impact on cancer research and therapy.</p><br />
</div></div>Amurthurhttp://2012.igem.org/Team:Penn/ProjectResultsTeam:Penn/ProjectResults2012-10-27T03:47:50Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">A Novel Therapeutic Platform</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">What if you could combine spatial targeting and cellular targeting into the same therapeutic? This idea is unprecedented but would allow for precise targeting of specific cells within a specific area, leaving healthy tissue intact and keeping side effects to a minimum. Higher dose precision means more of the therapeutic would be used efficiently in the targeted area and the dependency on passive diffusion – and the uncertainties that comes with it – would be eliminated.</p><br />
<br />
<p style="color:black;text-indent:30px;">The 2012 Penn iGEM team has engineered a novel, <b>modular</b> platform for targeted therapeutics that employs simultaneous spatial and cellular targeting. We have achieved spatial (and temporal) targeting with a blue light-switchable transgene expression system and cellular targeting through display of an antibody-mimetic protein on the surface of E. coli for the first time. Our platform also enables more precise dose control in the targeted area through the length of blue-light exposure, which allows us to regulate effective levels of transgene expression.</p><br />
<br />
<p style="color:black;text-indent:30px;">As a proof of concept, we applied our system to the treatment of cancer, a disease in which spatial and cellular targeting are of utmost importance. We displayed a high-affinity antibody-mimetic protein that targets Human Epidermal Growth Factor Receptor 2 (HER2), a protein commonly overexpressed in cancer cells, especially in breast cancer tumors. We combined this cellular targeting with a light-activated cytotoxic protein delivery system to successfully target and kill breast cancer cells.</p><br />
</div><br />
<br />
<div style="background-color:#01256e;" align="center"><br />
<iframe width="700" height="393" style="margin-left:10px;padding:10px; background-color:#000000;"src="http://www.youtube.com/embed/0fqLD4IJMJo" frameborder="1" allowfullscreen></iframe><br />
</div> <br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Modularity</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">The strength in our platform lies in its modularity. Both the light-induced transgene expression system and the surface display system work and exist independently of each other on two compatible plasmids. These plasmids can be modified to meet the needs of any synthetic biologist. </p><br />
<br />
<p style="color:black;text-indent:30px;">Any gene of interest can be cloned into the light-induced transgene expression system and will then be expressed in a light-dependent and spatially controlled manner. Any targeting protein can be cloned into our surface display platform to allow cellular targeting against any desired biomarker. These modular plasmids may then be co-transformed together to create a bacterial therapeutic for a desired disease.</p><br />
<br />
<p style="color:black;text-indent:30px;">These modular components could also be extended into applications other than medical therapeutics, such as biocatalysis, manufacturing, and alternative energy. </p><br />
<br />
<br />
<b><div class="name" align="center">Generic Components</div></b><br />
<br />
<div align="center"><br />
<table width="860" cellspacing="20" style="background-color:#d7dce1;"><br />
<tr><br />
<td width="410"><img src="https://static.igem.org/mediawiki/2012/e/e9/Lightactivationmodule.gif" width="400" height="300" /><br />
</td><br />
<td width="410"><img src="https://static.igem.org/mediawiki/2012/5/5c/Surfdispmodule.gif"width = "400" height = "300" /><br />
</td><br />
</tr><br />
<tr valign="top"><br />
<td ><br />
<p style="text-align:justify;"><b>Spatial Targeting:</b> Surgeons excise a tumor manually, without regard for cellular heterogeneity within and around the tumor area.</p><br />
</td><br />
<td><br />
<p style="text-align:justify;"><b>Cellular Targeting:</b> Monoclonal antibodies identify antigens on certain cells or viruses. Monoclonal antibodies are often coupled with therapeutic agents. However, if the antigen is present in healthy tissue outside the diseased area, it will be targeted as well.</p><br />
</td><br />
</tr><br />
<br />
</table></div><br />
</div><br />
<br />
<br />
<br />
<br />
<br />
<div class="bigbox"><br />
<br />
<br />
<br />
<br />
<b><div class="name" align="center">Proof of Concept</div></b><br />
<br><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/f/f9/Full_Color_Complete_Schematic.gif" width="700" height="500" /></div><br />
<p style="color:black;text-indent:30px;"> For our engineered bacterial therapeutic, we chose to target breast cancer as a proof of concept using a blue light gene expression system. We chose this specific blue-light inducible gene expression system because of the well-characterized nature of the parts and the high on:off ratio.</p><br />
<br />
<p style="color:black;text-indent:30px;"> We also identified a well-characterized class of antibody mimetic proteins called designed ankryin repeat proteins, or DARPins. One DARPin in particular (H10-2-G3) was engineered to bind to the Human Epidermal Growth Factor 2 (HER2) at picomolar affinities. HER2 is overexpressed in breast cancer cells, and we had access to cell lines that overexpressed HER2 on their cell surface which we could use for binding assays. </p><br />
<br />
</div><br />
<br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Human Practices</div></b><br />
<br><br />
<br />
<p style="color:black;text-indent:30px;">Upon conception of this project, we realized that although hundreds of academic research projects and iGEM projects have been conducted in the realm of Health and Medicine, almost no engineered bacterial therapeutics have been brought to the clinic. We analyzed the hurdles and road ahead for bacterial synthetic biology-enabled therapeutics, compiling a thorough report with specific actions that iGEM teams in Health/Medicine can take to make their therapies more clinically tractable. This project directly informed our wet lab work, leading us to port our therapeutic system into a non-pathogenic, probiotic bacterial strain which is already used in human therapies today.</p><br />
<br />
<br />
<p style="color:black;text-indent:30px;">We hope our targeted therapeutic platform will allow other scientists and iGEM teams to target any cells they choose. In the near term, we are planning to test our cancer cell targeting/killing bacterial system in a mouse model and make a real impact on cancer research and therapy.</p><br />
</div></div>Amurthurhttp://2012.igem.org/Team:Penn/ProjectResultsTeam:Penn/ProjectResults2012-10-27T03:43:24Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">A Novel Therapeutic Platform</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">What if you could combine spatial targeting and cellular targeting into the same therapeutic? This idea is unprecedented but would allow for precise targeting of specific cells within a specific area, leaving healthy tissue intact and keeping side effects to a minimum. Higher dose precision means more of the therapeutic would be used efficiently in the targeted area and the dependency on passive diffusion – and the uncertainties that comes with it – would be eliminated.</p><br />
<br />
<p style="color:black;text-indent:30px;">The 2012 Penn iGEM team has engineered a novel, <b>modular</b> platform for targeted therapeutics that employs simultaneous spatial and cellular targeting. We have achieved spatial (and temporal) targeting with a blue light-switchable transgene expression system and cellular targeting through display of an antibody-mimetic protein on the surface of E. coli for the first time. Our platform also enables more precise dose control in the targeted area through the length of blue-light exposure, which allows us to regulate effective levels of transgene expression.</p><br />
<br />
<p style="color:black;text-indent:30px;">As a proof of concept, we applied our system to the treatment of cancer, a disease in which spatial and cellular targeting are of utmost importance. We displayed a high-affinity antibody-mimetic protein that targets Human Epidermal Growth Factor Receptor 2 (HER2), a protein commonly overexpressed in cancer cells, especially in breast cancer tumors. We combined this cellular targeting with a light-activated cytotoxic protein delivery system to successfully target and kill breast cancer cells.</p><br />
</div><br />
<br />
<div style="background-color:#01256e;" align="center"><br />
<iframe width="700" height="393" style="margin-left:10px;padding:10px; background-color:#000000;"src="http://www.youtube.com/embed/0fqLD4IJMJo" frameborder="1" allowfullscreen></iframe><br />
</div> <br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Modularity</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">The strength in our platform lies in its modularity. Both the light-induced transgene expression system and the surface display system work and exist independently of each other on two compatible plasmids. These plasmids can be modified to meet the needs of any synthetic biologist. </p><br />
<br />
<p style="color:black;text-indent:30px;">Any gene of interest can be cloned into the light-induced transgene expression system and will then be expressed in a light-dependent and spatially controlled manner. Any targeting protein can be cloned into our surface display platform to allow cellular targeting against any desired biomarker. These modular plasmids may then be co-transformed together to create a bacterial therapeutic for a desired disease.</p><br />
<br />
<p style="color:black;text-indent:30px;">These modular components could also be extended into applications other than medical therapeutics, such as biocatalysis, manufacturing, and alternative energy. </p><br />
<br />
<br />
<b><div class="name" align="center">Generic Components</div></b><br />
<br />
<div align="center"><br />
<table width="860" cellspacing="20" style="background-color:#d7dce1;"><br />
<tr><br />
<td width="410"><img src="https://static.igem.org/mediawiki/2012/e/e9/Lightactivationmodule.gif" width="400" height="300" /><br />
</td><br />
<td width="410"><img src="https://static.igem.org/mediawiki/2012/5/5c/Surfdispmodule.gif"width = "400" height = "300" /><br />
</td><br />
</tr><br />
<tr valign="top"><br />
<td ><br />
<p style="text-align:justify;"><b>Spatial Targeting:</b> Surgeons excise a tumor manually, without regard for cellular heterogeneity within and around the tumor area.</p><br />
</td><br />
<td><br />
<p style="text-align:justify;"><b>Cellular Targeting:</b> Monoclonal antibodies identify antigens on certain cells or viruses. Monoclonal antibodies are often coupled with therapeutic agents. However, if the antigen is present in healthy tissue outside the diseased area, it will be targeted as well.</p><br />
</td><br />
</tr><br />
<br />
</table></div><br />
</div><br />
<br />
<br />
<br />
<br />
<br />
<div class="bigbox"><br />
<br />
<br />
<br />
<br />
<b><div class="name" align="center">Proof of Concept</div></b><br />
<br><br />
<br />
<p style="color:black;text-indent:30px;"> For our engineered bacterial therapeutic, we chose to target breast cancer as a proof of concept using a blue light gene expression system. We chose this specific blue-light inducible gene expression system because of the well-characterized nature of the parts and the high on:off ratio.</p><br />
<br />
<p style="color:black;text-indent:30px;"> We also identified a well-characterized class of antibody mimetic proteins called designed ankryin repeat proteins, or DARPins. One DARPin in particular (H10-2-G3) was engineered to bind to the Human Epidermal Growth Factor 2 (HER2) at picomolar affinities. HER2 is overexpressed in breast cancer cells, and we had access to cell lines that overexpressed HER2 on their cell surface which we could use for binding assays. </p><br />
<br />
</div><br />
<br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Human Practices</div></b><br />
<br><br />
<br />
<p style="color:black;text-indent:30px;">Upon conception of this project, we realized that although hundreds of academic research projects and iGEM projects have been conducted in the realm of Health and Medicine, almost no engineered bacterial therapeutics have been brought to the clinic. We analyzed the hurdles and road ahead for bacterial synthetic biology-enabled therapeutics, compiling a thorough report with specific actions that iGEM teams in Health/Medicine can take to make their therapies more clinically tractable. This project directly informed our wet lab work, leading us to port our therapeutic system into a non-pathogenic, probiotic bacterial strain which is already used in human therapies today.</p><br />
<br />
<br />
<p style="color:black;text-indent:30px;">We hope our targeted therapeutic platform will allow other scientists and iGEM teams to target any cells they choose. In the near term, we are planning to test our cancer cell targeting/killing bacterial system in a mouse model and make a real impact on cancer research and therapy.</p><br />
</div></div>Amurthurhttp://2012.igem.org/Team:Penn/ProjectResultsTeam:Penn/ProjectResults2012-10-27T03:42:00Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">A Novel Therapeutic Platform</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">What if you could combine spatial targeting and cellular targeting into the same therapeutic? This idea is unprecedented but would allow for precise targeting of specific cells within a specific area, leaving healthy tissue intact and keeping side effects to a minimum. Higher dose precision means more of the therapeutic would be used efficiently in the targeted area and the dependency on passive diffusion – and the uncertainties that comes with it – would be eliminated.</p><br />
<br />
<p style="color:black;text-indent:30px;">The 2012 Penn iGEM team has engineered a novel, <b>modular</b> platform for targeted therapeutics that employs simultaneous spatial and cellular targeting. We have achieved spatial (and temporal) targeting with a blue light-switchable transgene expression system and cellular targeting through display of an antibody-mimetic protein on the surface of E. coli for the first time. Our platform also enables more precise dose control in the targeted area through the length of blue-light exposure, which allows us to regulate effective levels of transgene expression.</p><br />
<br />
<p style="color:black;text-indent:30px;">As a proof of concept, we applied our system to the treatment of cancer, a disease in which spatial and cellular targeting are of utmost importance. We displayed a high-affinity antibody-mimetic protein that targets Human Epidermal Growth Factor Receptor 2 (HER2), a protein commonly overexpressed in cancer cells, especially in breast cancer tumors. We combined this cellular targeting with a light-activated cytotoxic protein delivery system to successfully target and kill breast cancer cells.</p><br />
</div><br />
<br />
<div style="background-color:#01256e;" align="center"><br />
<iframe width="700" height="393" style="margin-left:10px;padding:10px; background-color:#000000;"src="http://www.youtube.com/embed/0fqLD4IJMJo" frameborder="1" allowfullscreen></iframe><br />
</div> <br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Modularity</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">The strength in our platform lies in its modularity. Both the light-induced transgene expression system and the surface display system work and exist independently of each other on two compatible plasmids. These plasmids can be modified to meet the needs of any synthetic biologist. </p><br />
<br />
<p style="color:black;text-indent:30px;">Any gene of interest can be cloned into the light-induced transgene expression system and will then be expressed in a light-dependent and spatially controlled manner. Any targeting protein can be cloned into our surface display platform to allow cellular targeting against any desired biomarker. These modular plasmids may then be co-transformed together to create a bacterial therapeutic for a desired disease.</p><br />
<br />
<p style="color:black;text-indent:30px;">These modular components could also be extended into applications other than medical therapeutics, such as biocatalysis, manufacturing, and alternative energy. </p><br />
<br />
<br />
<b><div class="name" align="center">Generic Components</div></b><br />
<br />
<div align="center"><br />
<table width="860" cellspacing="20" style="background-color:#d7dce1;"><br />
<tr><br />
<td width="410"><img src="https://static.igem.org/mediawiki/2012/e/e9/Lightactivationmodule.gif" width="400" height="300" /><br />
</td><br />
<td width="410"><img src="https://static.igem.org/mediawiki/2012/6/6b/Cellular_Targeting.jpg" width = "400" height = "300" /><br />
</td><br />
</tr><br />
<tr valign="top"><br />
<td ><br />
<p style="text-align:justify;"><b>Spatial Targeting:</b> Surgeons excise a tumor manually, without regard for cellular heterogeneity within and around the tumor area.</p><br />
</td><br />
<td><br />
<p style="text-align:justify;"><b>Cellular Targeting:</b> Monoclonal antibodies identify antigens on certain cells or viruses. Monoclonal antibodies are often coupled with therapeutic agents. However, if the antigen is present in healthy tissue outside the diseased area, it will be targeted as well.</p><br />
</td><br />
</tr><br />
<br />
</table></div><br />
</div><br />
<br />
<br />
<br />
<br />
<br />
<div class="bigbox"><br />
<br />
<br />
<br />
<br />
<b><div class="name" align="center">Proof of Concept</div></b><br />
<br><br />
<br />
<p style="color:black;text-indent:30px;"> For our engineered bacterial therapeutic, we chose to target breast cancer as a proof of concept using a blue light gene expression system. We chose this specific blue-light inducible gene expression system because of the well-characterized nature of the parts and the high on:off ratio.</p><br />
<br />
<p style="color:black;text-indent:30px;"> We also identified a well-characterized class of antibody mimetic proteins called designed ankryin repeat proteins, or DARPins. One DARPin in particular (H10-2-G3) was engineered to bind to the Human Epidermal Growth Factor 2 (HER2) at picomolar affinities. HER2 is overexpressed in breast cancer cells, and we had access to cell lines that overexpressed HER2 on their cell surface which we could use for binding assays. </p><br />
<br />
</div><br />
<br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Human Practices</div></b><br />
<br><br />
<br />
<p style="color:black;text-indent:30px;">Upon conception of this project, we realized that although hundreds of academic research projects and iGEM projects have been conducted in the realm of Health and Medicine, almost no engineered bacterial therapeutics have been brought to the clinic. We analyzed the hurdles and road ahead for bacterial synthetic biology-enabled therapeutics, compiling a thorough report with specific actions that iGEM teams in Health/Medicine can take to make their therapies more clinically tractable. This project directly informed our wet lab work, leading us to port our therapeutic system into a non-pathogenic, probiotic bacterial strain which is already used in human therapies today.</p><br />
<br />
<br />
<p style="color:black;text-indent:30px;">We hope our targeted therapeutic platform will allow other scientists and iGEM teams to target any cells they choose. In the near term, we are planning to test our cancer cell targeting/killing bacterial system in a mouse model and make a real impact on cancer research and therapy.</p><br />
</div></div>Amurthurhttp://2012.igem.org/Team:Penn/ProjectResultsTeam:Penn/ProjectResults2012-10-27T03:41:42Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">A Novel Therapeutic Platform</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">What if you could combine spatial targeting and cellular targeting into the same therapeutic? This idea is unprecedented but would allow for precise targeting of specific cells within a specific area, leaving healthy tissue intact and keeping side effects to a minimum. Higher dose precision means more of the therapeutic would be used efficiently in the targeted area and the dependency on passive diffusion – and the uncertainties that comes with it – would be eliminated.</p><br />
<br />
<p style="color:black;text-indent:30px;">The 2012 Penn iGEM team has engineered a novel, <b>modular</b> platform for targeted therapeutics that employs simultaneous spatial and cellular targeting. We have achieved spatial (and temporal) targeting with a blue light-switchable transgene expression system and cellular targeting through display of an antibody-mimetic protein on the surface of E. coli for the first time. Our platform also enables more precise dose control in the targeted area through the length of blue-light exposure, which allows us to regulate effective levels of transgene expression.</p><br />
<br />
<p style="color:black;text-indent:30px;">As a proof of concept, we applied our system to the treatment of cancer, a disease in which spatial and cellular targeting are of utmost importance. We displayed a high-affinity antibody-mimetic protein that targets Human Epidermal Growth Factor Receptor 2 (HER2), a protein commonly overexpressed in cancer cells, especially in breast cancer tumors. We combined this cellular targeting with a light-activated cytotoxic protein delivery system to successfully target and kill breast cancer cells.</p><br />
</div><br />
<br />
<div style="background-color:#01256e;" align="center"><br />
<iframe width="700" height="393" style="margin-left:10px;padding:10px; background-color:#000000;"src="http://www.youtube.com/embed/0fqLD4IJMJo" frameborder="1" allowfullscreen></iframe><br />
</div> <br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Modularity</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">The strength in our platform lies in its modularity. Both the light-induced transgene expression system and the surface display system work and exist independently of each other on two compatible plasmids. These plasmids can be modified to meet the needs of any synthetic biologist. </p><br />
<br />
<p style="color:black;text-indent:30px;">Any gene of interest can be cloned into the light-induced transgene expression system and will then be expressed in a light-dependent and spatially controlled manner. Any targeting protein can be cloned into our surface display platform to allow cellular targeting against any desired biomarker. These modular plasmids may then be co-transformed together to create a bacterial therapeutic for a desired disease.</p><br />
<br />
<p style="color:black;text-indent:30px;">These modular components could also be extended into applications other than medical therapeutics, such as biocatalysis, manufacturing, and alternative energy. </p><br />
<br />
<br />
<b><div class="name" align="center">Generic Components</div></b><br />
<br />
<div align="center"><br />
<table width="860" cellspacing="20" style="background-color:#d7dce1;"><br />
<tr><br />
<td width="410"><img src="https://static.igem.org/mediawiki/2012/f/fa/Spatial_Targeting.jpg" width="400" height="300" /><br />
</td><br />
<td width="410"><img src="https://static.igem.org/mediawiki/2012/6/6b/Cellular_Targeting.jpg" width = "400" height = "300" /><br />
</td><br />
</tr><br />
<tr valign="top"><br />
<td ><br />
<p style="text-align:justify;"><b>Spatial Targeting:</b> Surgeons excise a tumor manually, without regard for cellular heterogeneity within and around the tumor area.</p><br />
</td><br />
<td><br />
<p style="text-align:justify;"><b>Cellular Targeting:</b> Monoclonal antibodies identify antigens on certain cells or viruses. Monoclonal antibodies are often coupled with therapeutic agents. However, if the antigen is present in healthy tissue outside the diseased area, it will be targeted as well.</p><br />
</td><br />
</tr><br />
<br />
</table></div><br />
</div><br />
<br />
<br />
<br />
<br />
<br />
<div class="bigbox"><br />
<br />
<br />
<br />
<br />
<b><div class="name" align="center">Proof of Concept</div></b><br />
<br><br />
<br />
<p style="color:black;text-indent:30px;"> For our engineered bacterial therapeutic, we chose to target breast cancer as a proof of concept using a blue light gene expression system. We chose this specific blue-light inducible gene expression system because of the well-characterized nature of the parts and the high on:off ratio.</p><br />
<br />
<p style="color:black;text-indent:30px;"> We also identified a well-characterized class of antibody mimetic proteins called designed ankryin repeat proteins, or DARPins. One DARPin in particular (H10-2-G3) was engineered to bind to the Human Epidermal Growth Factor 2 (HER2) at picomolar affinities. HER2 is overexpressed in breast cancer cells, and we had access to cell lines that overexpressed HER2 on their cell surface which we could use for binding assays. </p><br />
<br />
</div><br />
<br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Human Practices</div></b><br />
<br><br />
<br />
<p style="color:black;text-indent:30px;">Upon conception of this project, we realized that although hundreds of academic research projects and iGEM projects have been conducted in the realm of Health and Medicine, almost no engineered bacterial therapeutics have been brought to the clinic. We analyzed the hurdles and road ahead for bacterial synthetic biology-enabled therapeutics, compiling a thorough report with specific actions that iGEM teams in Health/Medicine can take to make their therapies more clinically tractable. This project directly informed our wet lab work, leading us to port our therapeutic system into a non-pathogenic, probiotic bacterial strain which is already used in human therapies today.</p><br />
<br />
<br />
<p style="color:black;text-indent:30px;">We hope our targeted therapeutic platform will allow other scientists and iGEM teams to target any cells they choose. In the near term, we are planning to test our cancer cell targeting/killing bacterial system in a mouse model and make a real impact on cancer research and therapy.</p><br />
</div></div>Amurthurhttp://2012.igem.org/Team:Penn/ProjectResultsTeam:Penn/ProjectResults2012-10-27T03:40:56Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">A Novel Therapeutic Platform</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">What if you could combine spatial targeting and cellular targeting into the same therapeutic? This idea is unprecedented but would allow for precise targeting of specific cells within a specific area, leaving healthy tissue intact and keeping side effects to a minimum. Higher dose precision means more of the therapeutic would be used efficiently in the targeted area and the dependency on passive diffusion – and the uncertainties that comes with it – would be eliminated.</p><br />
<br />
<p style="color:black;text-indent:30px;">The 2012 Penn iGEM team has engineered a novel, <b>modular</b> platform for targeted therapeutics that employs simultaneous spatial and cellular targeting. We have achieved spatial (and temporal) targeting with a blue light-switchable transgene expression system and cellular targeting through display of an antibody-mimetic protein on the surface of E. coli for the first time. Our platform also enables more precise dose control in the targeted area through the length of blue-light exposure, which allows us to regulate effective levels of transgene expression.</p><br />
<br />
<p style="color:black;text-indent:30px;">As a proof of concept, we applied our system to the treatment of cancer, a disease in which spatial and cellular targeting are of utmost importance. We displayed a high-affinity antibody-mimetic protein that targets Human Epidermal Growth Factor Receptor 2 (HER2), a protein commonly overexpressed in cancer cells, especially in breast cancer tumors. We combined this cellular targeting with a light-activated cytotoxic protein delivery system to successfully target and kill breast cancer cells.</p><br />
</div><br />
<br />
<div style="background-color:#01256e;" align="center"><br />
<iframe width="700" height="393" style="margin-left:10px;padding:10px; background-color:#000000;"src="http://www.youtube.com/embed/0fqLD4IJMJo" frameborder="1" allowfullscreen></iframe><br />
</div> <br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Modularity</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">The strength in our platform lies in its modularity. Both the light-induced transgene expression system and the surface display system work and exist independently of each other on two compatible plasmids. These plasmids can be modified to meet the needs of any synthetic biologist. </p><br />
<br />
<p style="color:black;text-indent:30px;">Any gene of interest can be cloned into the light-induced transgene expression system and will then be expressed in a light-dependent and spatially controlled manner. Any targeting protein can be cloned into our surface display platform to allow cellular targeting against any desired biomarker. These modular plasmids may then be co-transformed together to create a bacterial therapeutic for a desired disease.</p><br />
<br />
<p style="color:black;text-indent:30px;">These modular components could also be extended into applications other than medical therapeutics, such as biocatalysis, manufacturing, and alternative energy. </p><br />
<br />
<br />
<b><div class="name" align="center">Generic Components</div></b><br />
<br />
<div align="center"><br />
<table width="860" cellspacing="20" style="background-color:#d7dce1;"><br />
<tr><br />
<td width="410"><img src="https://static.igem.org/mediawiki/2012/f/fa/Spatial_Targeting.jpg" width="400" height="300" /><br />
</td><br />
<td width="410"><img src="https://static.igem.org/mediawiki/2012/6/6b/Cellular_Targeting.jpg" width = "400" height = "300" /><br />
</td><br />
</tr><br />
<tr valign="top"><br />
<td ><br />
<p style="text-align:justify;"><b>Spatial Targeting:</b> Surgeons excise a tumor manually, without regard for cellular heterogeneity within and around the tumor area.</p><br />
</td><br />
<td><br />
<p style="text-align:justify;"><b>Cellular Targeting:</b> Monoclonal antibodies identify antigens on certain cells or viruses. Monoclonal antibodies are often coupled with therapeutic agents. However, if the antigen is present in healthy tissue outside the diseased area, it will be targeted as well.</p><br />
</td><br />
</tr><br />
<br />
</table></div><br />
<br />
<br />
<br />
<br />
<br />
<br />
<div class="bigbox"><br />
<br />
<br />
<br />
<br />
<b><div class="name" align="center">Proof of Concept</div></b><br />
<br><br />
<br />
<p style="color:black;text-indent:30px;"> For our engineered bacterial therapeutic, we chose to target breast cancer as a proof of concept using a blue light gene expression system. We chose this specific blue-light inducible gene expression system because of the well-characterized nature of the parts and the high on:off ratio.</p><br />
<br />
<p style="color:black;text-indent:30px;"> We also identified a well-characterized class of antibody mimetic proteins called designed ankryin repeat proteins, or DARPins. One DARPin in particular (H10-2-G3) was engineered to bind to the Human Epidermal Growth Factor 2 (HER2) at picomolar affinities. HER2 is overexpressed in breast cancer cells, and we had access to cell lines that overexpressed HER2 on their cell surface which we could use for binding assays. </p><br />
<br />
</div><br />
<br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Human Practices</div></b><br />
<br><br />
<br />
<p style="color:black;text-indent:30px;">Upon conception of this project, we realized that although hundreds of academic research projects and iGEM projects have been conducted in the realm of Health and Medicine, almost no engineered bacterial therapeutics have been brought to the clinic. We analyzed the hurdles and road ahead for bacterial synthetic biology-enabled therapeutics, compiling a thorough report with specific actions that iGEM teams in Health/Medicine can take to make their therapies more clinically tractable. This project directly informed our wet lab work, leading us to port our therapeutic system into a non-pathogenic, probiotic bacterial strain which is already used in human therapies today.</p><br />
<br />
<br />
<p style="color:black;text-indent:30px;">We hope our targeted therapeutic platform will allow other scientists and iGEM teams to target any cells they choose. In the near term, we are planning to test our cancer cell targeting/killing bacterial system in a mouse model and make a real impact on cancer research and therapy.</p><br />
</div></div>Amurthurhttp://2012.igem.org/Team:Penn/ProjectResultsTeam:Penn/ProjectResults2012-10-27T03:38:35Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">A Novel Therapeutic Platform</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">What if you could combine spatial targeting and cellular targeting into the same therapeutic? This idea is unprecedented but would allow for precise targeting of specific cells within a specific area, leaving healthy tissue intact and keeping side effects to a minimum. Higher dose precision means more of the therapeutic would be used efficiently in the targeted area and the dependency on passive diffusion – and the uncertainties that comes with it – would be eliminated.</p><br />
<br />
<p style="color:black;text-indent:30px;">The 2012 Penn iGEM team has engineered a novel, <b>modular</b> platform for targeted therapeutics that employs simultaneous spatial and cellular targeting. We have achieved spatial (and temporal) targeting with a blue light-switchable transgene expression system and cellular targeting through display of an antibody-mimetic protein on the surface of E. coli for the first time. Our platform also enables more precise dose control in the targeted area through the length of blue-light exposure, which allows us to regulate effective levels of transgene expression.</p><br />
<br />
<p style="color:black;text-indent:30px;">As a proof of concept, we applied our system to the treatment of cancer, a disease in which spatial and cellular targeting are of utmost importance. We displayed a high-affinity antibody-mimetic protein that targets Human Epidermal Growth Factor Receptor 2 (HER2), a protein commonly overexpressed in cancer cells, especially in breast cancer tumors. We combined this cellular targeting with a light-activated cytotoxic protein delivery system to successfully target and kill breast cancer cells.</p><br />
</div><br />
<br />
<div style="background-color:#01256e;" align="center"><br />
<iframe width="700" height="393" style="margin-left:10px;padding:10px; background-color:#000000;"src="http://www.youtube.com/embed/0fqLD4IJMJo" frameborder="1" allowfullscreen></iframe><br />
</div> <br><br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Modularity</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">The strength in our platform lies in its modularity. Both the light-induced transgene expression system and the surface display system work and exist independently of each other on two compatible plasmids. These plasmids can be modified to meet the needs of any synthetic biologist. </p><br />
<br />
<p style="color:black;text-indent:30px;">Any gene of interest can be cloned into the light-induced transgene expression system and will then be expressed in a light-dependent and spatially controlled manner. Any targeting protein can be cloned into our surface display platform to allow cellular targeting against any desired biomarker. These modular plasmids may then be co-transformed together to create a bacterial therapeutic for a desired disease.</p><br />
<br />
<p style="color:black;text-indent:30px;">These modular components could also be extended into applications other than medical therapeutics, such as biocatalysis, manufacturing, and alternative energy. </p><br />
</div><br />
<br />
<b><div class="name" align="center">Generic Components</div></b><br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<div class="bigbox"><br />
<br />
<br />
<br />
<br />
<b><div class="name" align="center">Proof of Concept</div></b><br />
<br><br />
<br />
</div><br />
<br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Human Practices</div></b><br />
<br><br />
<br />
<p style="color:black;text-indent:30px;">Upon conception of this project, we realized that although hundreds of academic research projects and iGEM projects have been conducted in the realm of Health and Medicine, almost no engineered bacterial therapeutics have been brought to the clinic. We analyzed the hurdles and road ahead for bacterial synthetic biology-enabled therapeutics, compiling a thorough report with specific actions that iGEM teams in Health/Medicine can take to make their therapies more clinically tractable. This project directly informed our wet lab work, leading us to port our therapeutic system into a non-pathogenic, probiotic bacterial strain which is already used in human therapies today.</p><br />
<br />
<br />
<p style="color:black;text-indent:30px;">We hope our targeted therapeutic platform will allow other scientists and iGEM teams to target any cells they choose. In the near term, we are planning to test our cancer cell targeting/killing bacterial system in a mouse model and make a real impact on cancer research and therapy.</p><br />
</div></div>Amurthurhttp://2012.igem.org/Team:Penn/SurfaceDisplayOverviewTeam:Penn/SurfaceDisplayOverview2012-10-27T03:33:36Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
.figs2{width:916px; margin:0 auto; overflow:hidden;}<br />
.fignew{font-size:13px; width:418px; margin:10px auto; float:left; padding:0 20px 0 20px;}<br />
.fig{font-size:13px; width:500px; margin:10px auto;}<br />
</style><br />
<br><br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Surface Display and Targeting</div></b><br><br />
<br />
<div align="center"><img style="align:center;"src="https://static.igem.org/mediawiki/2012/a/a4/Surfacedispschematic.gif" height="525" width="700"></div><br><br />
While engineering bacteria as a light-activated drug delivery <a href="https://2012.igem.org/Team:Penn/LightActivatedOverview "> system </a> is novel and useful by itself, the targeting potential of our light-activated bacterial therapeutic would be greatly improved if the bacteria also could target specific cells. The goal of the second module of our system is to allow bacterial targeting to cancer cells. We sought to achieve this by displaying an engineered cancer cell binding protein on the surface of our <i> E. coli</i>. We successfully displayed DARPin H10-2-G3, an antibody-mimetic protein rationally evolved to picomolar affinity with HER2, a breast cancer biomarker. We verified that bacteria displaying this protein can selectively bind to breast cancer cells <i> in vitro</i>. This class of protein has not been displayed before on the surface of bacteria. Along the way, we developed a generalized BioBrick surface display system which allows future iGEM teams to display proteins of their choosing on the surface of bacteria.<br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objectives</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">Our goals were twofold:<br />
<ol style="font-size:15px"><li><b> Achieve the first display of DARPin H10-2-G3 on the surface of <i> E. coli</i> and verify that the system targets cancer cells. </b></li><br />
<li><b> Create a generalized BioBrick surface display platform for other labs and iGEM teams.</b></li><br />
</ul><br />
</div><br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Ice Nucleation Protein</div></b><br />
<br><br />
We chose to use the Ice Nucleation Protein (Figure 1) as our surface display carrier protein. The Ice Nucleation Protein protein has been used to display enzymes [1], typically for biocatalysis applications (e.g. Edinburgh iGEM 2011). To make the protein a more manageable size, we truncated the protein to the N and C terminal domains only. The C terminal domain is displayed at the cell surface, while the N terminal domain remains in the outer membrane. We sought to apply this system to health/medicine by displaying DARPin H10-2-G3.<br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/e/e4/INPNC.png" /><br><br />
<b>Figure 1</b></div><div style="text-align:center">Figure 1: Proposed Structure of Ice Nucleation Protein.</div></div><br />
<p style="color:black"><br><br><br />
[1] Zahnd, C., Wyler, E., Schwenk, J. M., Steiner, D., Lawrence, M. C., McKern, N. M., Pecorari, F., et al. (2007). A designed ankyrin repeat protein evolved to picomolar affinity to Her2. Journal of molecular biology, 369(4), 1015–28. doi:10.1016/j.jmb.2007.03.028 <br />
</p><br />
</div><br />
<br><br />
<br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">DARPin H10-2-G3</div></b><br />
<br><br />
The <a href="http://www.bioc.uzh.ch/plueckthun/"> lab </a> of Andreas Plueckthun at ETH Zurich has pioneered the development of Designed Ankyrin Repeat Protein (DARPin) technology. DARPins are engineered antibody-mimetic proteins consisting of 3-5 ankyrin repeat motifs. These proteins have been rationally evolved to high binding affinity with targets through ribosome display. In 2007, the group developed H10-2-G3 [2], a 14.7kDa DARPin evolved to 90pM affinity with the extracellular domain of HER2 (Figure 2).<br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/b/b7/DARPingif.gif" /><br><br />
<b>Figure 2</b></div><div style="text-align:center">Figure 2: Structure of DARPin-H10-2-G3.</div></div><br />
<p style="color:black"><br><br><br />
[2] Zahnd, C., Wyler, E., Schwenk, J. M., Steiner, D., Lawrence, M. C., McKern, N. M., Pecorari, F., et al. (2007). A designed ankyrin repeat protein evolved to picomolar affinity to Her2. Journal of molecular biology, 369(4), 1015–28. doi:10.1016/j.jmb.2007.03.028 <br />
</p><br />
</div><br />
<br><br />
<br />
<br />
<br />
<br />
<br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/SurfaceDisplayOverviewTeam:Penn/SurfaceDisplayOverview2012-10-27T03:33:26Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
.figs2{width:916px; margin:0 auto; overflow:hidden;}<br />
.fignew{font-size:13px; width:418px; margin:10px auto; float:left; padding:0 20px 0 20px;}<br />
.fig{font-size:13px; width:500px; margin:10px auto;}<br />
</style><br />
<br><br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Surface Display and Targeting</div></b><br><br />
<br />
<div align="center"><img style="align:center;"src="https://static.igem.org/mediawiki/2012/a/a4/Surfacedispschematic.gif" height="525" width="700"></div><br />
While engineering bacteria as a light-activated drug delivery <a href="https://2012.igem.org/Team:Penn/LightActivatedOverview "> system </a> is novel and useful by itself, the targeting potential of our light-activated bacterial therapeutic would be greatly improved if the bacteria also could target specific cells. The goal of the second module of our system is to allow bacterial targeting to cancer cells. We sought to achieve this by displaying an engineered cancer cell binding protein on the surface of our <i> E. coli</i>. We successfully displayed DARPin H10-2-G3, an antibody-mimetic protein rationally evolved to picomolar affinity with HER2, a breast cancer biomarker. We verified that bacteria displaying this protein can selectively bind to breast cancer cells <i> in vitro</i>. This class of protein has not been displayed before on the surface of bacteria. Along the way, we developed a generalized BioBrick surface display system which allows future iGEM teams to display proteins of their choosing on the surface of bacteria.<br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objectives</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">Our goals were twofold:<br />
<ol style="font-size:15px"><li><b> Achieve the first display of DARPin H10-2-G3 on the surface of <i> E. coli</i> and verify that the system targets cancer cells. </b></li><br />
<li><b> Create a generalized BioBrick surface display platform for other labs and iGEM teams.</b></li><br />
</ul><br />
</div><br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Ice Nucleation Protein</div></b><br />
<br><br />
We chose to use the Ice Nucleation Protein (Figure 1) as our surface display carrier protein. The Ice Nucleation Protein protein has been used to display enzymes [1], typically for biocatalysis applications (e.g. Edinburgh iGEM 2011). To make the protein a more manageable size, we truncated the protein to the N and C terminal domains only. The C terminal domain is displayed at the cell surface, while the N terminal domain remains in the outer membrane. We sought to apply this system to health/medicine by displaying DARPin H10-2-G3.<br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/e/e4/INPNC.png" /><br><br />
<b>Figure 1</b></div><div style="text-align:center">Figure 1: Proposed Structure of Ice Nucleation Protein.</div></div><br />
<p style="color:black"><br><br><br />
[1] Zahnd, C., Wyler, E., Schwenk, J. M., Steiner, D., Lawrence, M. C., McKern, N. M., Pecorari, F., et al. (2007). A designed ankyrin repeat protein evolved to picomolar affinity to Her2. Journal of molecular biology, 369(4), 1015–28. doi:10.1016/j.jmb.2007.03.028 <br />
</p><br />
</div><br />
<br><br />
<br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">DARPin H10-2-G3</div></b><br />
<br><br />
The <a href="http://www.bioc.uzh.ch/plueckthun/"> lab </a> of Andreas Plueckthun at ETH Zurich has pioneered the development of Designed Ankyrin Repeat Protein (DARPin) technology. DARPins are engineered antibody-mimetic proteins consisting of 3-5 ankyrin repeat motifs. These proteins have been rationally evolved to high binding affinity with targets through ribosome display. In 2007, the group developed H10-2-G3 [2], a 14.7kDa DARPin evolved to 90pM affinity with the extracellular domain of HER2 (Figure 2).<br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/b/b7/DARPingif.gif" /><br><br />
<b>Figure 2</b></div><div style="text-align:center">Figure 2: Structure of DARPin-H10-2-G3.</div></div><br />
<p style="color:black"><br><br><br />
[2] Zahnd, C., Wyler, E., Schwenk, J. M., Steiner, D., Lawrence, M. C., McKern, N. M., Pecorari, F., et al. (2007). A designed ankyrin repeat protein evolved to picomolar affinity to Her2. Journal of molecular biology, 369(4), 1015–28. doi:10.1016/j.jmb.2007.03.028 <br />
</p><br />
</div><br />
<br><br />
<br />
<br />
<br />
<br />
<br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/LightActivatedOverviewTeam:Penn/LightActivatedOverview2012-10-27T03:32:40Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
.figs2{width:916px; margin:0 auto; overflow:hidden;}<br />
.fignew{font-size:13px; width:418px; margin:10px auto; float:left; padding:0 20px 0 20px;}<br />
.fig{font-size:13px; width:500px; margin:10px auto;}<br />
</style><br />
<br><br />
<br />
<div style="text-align:center;font-size:34px;color:white;"><b>Light-Activated Cell Lysis </b></div> <br />
<br><br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Objectives</div></b><br><br />
<br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/f/fc/Lightdispschematic.gif" height="525" width="700" /></div><br />
<br><br />
<p style="color:black;text-indent:30px;">In order to develop a module for light activated cell lysis, we had to implement two elements:<br />
<ol style="font-size:15px"><li><b>Construct a light-activation system that can express a downstream gene of interest.</b></li><br />
<li><b>Express a cytolytic protein that can be expressed as our therapeutic drug to lyse cancer cells.</b></li><br />
</ul><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 1: Light-Activated Sensor</div></b><br><br><br />
<b><div class="name" align="center" style="font-size:16px;">Selection of YF1/FixJ Blue Light Sensor</div></b><br><br />
<p style="color:black;text-indent:30px;">After reading many papers to select an appropriate light-sensing system to use, we selected the YF1/FixJ blue light system. We had also considered the red light sensor Cph8 but ultimately decided on YF1/FixJ because of its high on/off ratio of gene expression and also because of its availability to us (we were fortunate enough to come across the YF1/FixJ system in the form of the pDawn plasmid from the Moglich lab in Germany).</p><br><br />
<b><div class="name" align="center" style="font-size:16px;">YF1/FixJ System (pDawn)</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">As shown below in Figure 1, the YF1/FixJ system works through a "repress the repressor" concept. Upon 480 nm blue light illumination, YF1 (a fusion of a LOV protein domain and a histidine kinase) phosphorylates a FixJ response regulator that activates the pFixK2 promoter. The activation of pFixK2, promotes expression of the cI repressor that, in turn, represses the lambda promoter pR. The net result is activation of the gene in the downstream MCS. </p><br><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/7/74/PDAWN.gif" /><br />
</div><br />
<br><br />
<div style="text-align:center"><b>Figure 1</b><br /></div><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 2: Expression of a Cytolytic Protein</div></b><br><br />
<b><div class="name" align="center">Cytolysin A (ClyA)</div></b><br><br />
<p style="color:black;text-indent:30px;"><br />
ClyA is a protein native to E. coli, Shigella flexneri, and Salmonella typhi that is capable of forming 13-mer pore complexes in a redox-independent manner. Expression of clyA in the absence of other hemolytic toxins is sufficient to induce hemolysis experimentally, and is therefore considered to be a potent cytolytic agent. Unlike a similar protein, HlyA, ClyA is not synthesized as a protoxin, which requires further posttranslational modifications to become active. ClyA is functional immediately following translation of mRNA to protein.<br />
<br />
ClyA is a 34kDa protein that is composed primarily of α-helical bundles that form a rod-shaped molecule. The membrane insertion domain is known as a β tongue (shown in yellow in Figure 2) and is critical for hemolytic activity. If the β tongue is mutated, the hemolytic activity of clyA is abrogated. </p><br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/2/22/ClyAPoregif.gif" /><br><br />
<br><b>Figure 2</b></div>Figure 2: ClyA forms a 13-mer pore complex that consists of hydrophobic beta tongues (yellow) on the head domains of individual monomer units that play an important role in influencing its cytolytic functions.<br />
</div><br />
<br />
<br><br />
<b><div class="name" align="center">Mechanism of Action</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">We selected ClyA as our cytolytic protein because of its unique mechanism of action that makes it especially potent. ClyA is secreted from bacteria in outer membrane vesicles (OMV's), within which it forms pore assemblies. As shown below in Figure 3, these pore assemblies allow ClyA to latch on to the cell wall of other cells and through the use of its encapsulating pore assembly lyse the cell wall. </p><br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/6/6c/ClyA-Pore-Assembly.jpg" /><br><br />
<br><b>Figure 3</b></div>Figure 3: Shown above in the first image are pore assemblies containing 13-mers of ClyA interacting with the surface of the target membrane. The second image below shows the ClyA assembly lysing the cell membrane through pore formation. (Wallace et. al 2000)<br />
</div><br />
<br />
<br />
<br />
</body><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/LightActivatedOverviewTeam:Penn/LightActivatedOverview2012-10-27T03:32:00Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
.figs2{width:916px; margin:0 auto; overflow:hidden;}<br />
.fignew{font-size:13px; width:418px; margin:10px auto; float:left; padding:0 20px 0 20px;}<br />
.fig{font-size:13px; width:500px; margin:10px auto;}<br />
</style><br />
<br><br />
<br />
<div style="text-align:center;font-size:34px;color:white;"><b>Light-Activated Cell Lysis </b></div> <br />
<br><br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Objectives</div></b><br><br />
<br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/f/fc/Lightdispschematic.gif" height="500" width="700" /></div><br />
<br><br />
<p style="color:black;text-indent:30px;">In order to develop a module for light activated cell lysis, we had to implement two elements:<br />
<ol style="font-size:15px"><li><b>Construct a light-activation system that can express a downstream gene of interest.</b></li><br />
<li><b>Express a cytolytic protein that can be expressed as our therapeutic drug to lyse cancer cells.</b></li><br />
</ul><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 1: Light-Activated Sensor</div></b><br><br><br />
<b><div class="name" align="center" style="font-size:16px;">Selection of YF1/FixJ Blue Light Sensor</div></b><br><br />
<p style="color:black;text-indent:30px;">After reading many papers to select an appropriate light-sensing system to use, we selected the YF1/FixJ blue light system. We had also considered the red light sensor Cph8 but ultimately decided on YF1/FixJ because of its high on/off ratio of gene expression and also because of its availability to us (we were fortunate enough to come across the YF1/FixJ system in the form of the pDawn plasmid from the Moglich lab in Germany).</p><br><br />
<b><div class="name" align="center" style="font-size:16px;">YF1/FixJ System (pDawn)</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">As shown below in Figure 1, the YF1/FixJ system works through a "repress the repressor" concept. Upon 480 nm blue light illumination, YF1 (a fusion of a LOV protein domain and a histidine kinase) phosphorylates a FixJ response regulator that activates the pFixK2 promoter. The activation of pFixK2, promotes expression of the cI repressor that, in turn, represses the lambda promoter pR. The net result is activation of the gene in the downstream MCS. </p><br><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/7/74/PDAWN.gif" /><br />
</div><br />
<br><br />
<div style="text-align:center"><b>Figure 1</b><br /></div><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 2: Expression of a Cytolytic Protein</div></b><br><br />
<b><div class="name" align="center">Cytolysin A (ClyA)</div></b><br><br />
<p style="color:black;text-indent:30px;"><br />
ClyA is a protein native to E. coli, Shigella flexneri, and Salmonella typhi that is capable of forming 13-mer pore complexes in a redox-independent manner. Expression of clyA in the absence of other hemolytic toxins is sufficient to induce hemolysis experimentally, and is therefore considered to be a potent cytolytic agent. Unlike a similar protein, HlyA, ClyA is not synthesized as a protoxin, which requires further posttranslational modifications to become active. ClyA is functional immediately following translation of mRNA to protein.<br />
<br />
ClyA is a 34kDa protein that is composed primarily of α-helical bundles that form a rod-shaped molecule. The membrane insertion domain is known as a β tongue (shown in yellow in Figure 2) and is critical for hemolytic activity. If the β tongue is mutated, the hemolytic activity of clyA is abrogated. </p><br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/2/22/ClyAPoregif.gif" /><br><br />
<br><b>Figure 2</b></div>Figure 2: ClyA forms a 13-mer pore complex that consists of hydrophobic beta tongues (yellow) on the head domains of individual monomer units that play an important role in influencing its cytolytic functions.<br />
</div><br />
<br />
<br><br />
<b><div class="name" align="center">Mechanism of Action</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">We selected ClyA as our cytolytic protein because of its unique mechanism of action that makes it especially potent. ClyA is secreted from bacteria in outer membrane vesicles (OMV's), within which it forms pore assemblies. As shown below in Figure 3, these pore assemblies allow ClyA to latch on to the cell wall of other cells and through the use of its encapsulating pore assembly lyse the cell wall. </p><br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/6/6c/ClyA-Pore-Assembly.jpg" /><br><br />
<br><b>Figure 3</b></div>Figure 3: Shown above in the first image are pore assemblies containing 13-mers of ClyA interacting with the surface of the target membrane. The second image below shows the ClyA assembly lysing the cell membrane through pore formation. (Wallace et. al 2000)<br />
</div><br />
<br />
<br />
<br />
</body><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/LightActivatedOverviewTeam:Penn/LightActivatedOverview2012-10-27T03:31:52Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
.figs2{width:916px; margin:0 auto; overflow:hidden;}<br />
.fignew{font-size:13px; width:418px; margin:10px auto; float:left; padding:0 20px 0 20px;}<br />
.fig{font-size:13px; width:500px; margin:10px auto;}<br />
</style><br />
<br><br />
<br />
<div style="text-align:center;font-size:34px;color:white;"><b>Light-Activated Cell Lysis </b></div> <br />
<br><br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Objectives</div></b><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/f/fc/Lightdispschematic.gif" height="500" width="700" /></div><br />
<br><br />
<p style="color:black;text-indent:30px;">In order to develop a module for light activated cell lysis, we had to implement two elements:<br />
<ol style="font-size:15px"><li><b>Construct a light-activation system that can express a downstream gene of interest.</b></li><br />
<li><b>Express a cytolytic protein that can be expressed as our therapeutic drug to lyse cancer cells.</b></li><br />
</ul><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 1: Light-Activated Sensor</div></b><br><br><br />
<b><div class="name" align="center" style="font-size:16px;">Selection of YF1/FixJ Blue Light Sensor</div></b><br><br />
<p style="color:black;text-indent:30px;">After reading many papers to select an appropriate light-sensing system to use, we selected the YF1/FixJ blue light system. We had also considered the red light sensor Cph8 but ultimately decided on YF1/FixJ because of its high on/off ratio of gene expression and also because of its availability to us (we were fortunate enough to come across the YF1/FixJ system in the form of the pDawn plasmid from the Moglich lab in Germany).</p><br><br />
<b><div class="name" align="center" style="font-size:16px;">YF1/FixJ System (pDawn)</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">As shown below in Figure 1, the YF1/FixJ system works through a "repress the repressor" concept. Upon 480 nm blue light illumination, YF1 (a fusion of a LOV protein domain and a histidine kinase) phosphorylates a FixJ response regulator that activates the pFixK2 promoter. The activation of pFixK2, promotes expression of the cI repressor that, in turn, represses the lambda promoter pR. The net result is activation of the gene in the downstream MCS. </p><br><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/7/74/PDAWN.gif" /><br />
</div><br />
<br><br />
<div style="text-align:center"><b>Figure 1</b><br /></div><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 2: Expression of a Cytolytic Protein</div></b><br><br />
<b><div class="name" align="center">Cytolysin A (ClyA)</div></b><br><br />
<p style="color:black;text-indent:30px;"><br />
ClyA is a protein native to E. coli, Shigella flexneri, and Salmonella typhi that is capable of forming 13-mer pore complexes in a redox-independent manner. Expression of clyA in the absence of other hemolytic toxins is sufficient to induce hemolysis experimentally, and is therefore considered to be a potent cytolytic agent. Unlike a similar protein, HlyA, ClyA is not synthesized as a protoxin, which requires further posttranslational modifications to become active. ClyA is functional immediately following translation of mRNA to protein.<br />
<br />
ClyA is a 34kDa protein that is composed primarily of α-helical bundles that form a rod-shaped molecule. The membrane insertion domain is known as a β tongue (shown in yellow in Figure 2) and is critical for hemolytic activity. If the β tongue is mutated, the hemolytic activity of clyA is abrogated. </p><br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/2/22/ClyAPoregif.gif" /><br><br />
<br><b>Figure 2</b></div>Figure 2: ClyA forms a 13-mer pore complex that consists of hydrophobic beta tongues (yellow) on the head domains of individual monomer units that play an important role in influencing its cytolytic functions.<br />
</div><br />
<br />
<br><br />
<b><div class="name" align="center">Mechanism of Action</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">We selected ClyA as our cytolytic protein because of its unique mechanism of action that makes it especially potent. ClyA is secreted from bacteria in outer membrane vesicles (OMV's), within which it forms pore assemblies. As shown below in Figure 3, these pore assemblies allow ClyA to latch on to the cell wall of other cells and through the use of its encapsulating pore assembly lyse the cell wall. </p><br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/6/6c/ClyA-Pore-Assembly.jpg" /><br><br />
<br><b>Figure 3</b></div>Figure 3: Shown above in the first image are pore assemblies containing 13-mers of ClyA interacting with the surface of the target membrane. The second image below shows the ClyA assembly lysing the cell membrane through pore formation. (Wallace et. al 2000)<br />
</div><br />
<br />
<br />
<br />
</body><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/LightActivatedOverviewTeam:Penn/LightActivatedOverview2012-10-27T03:31:36Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
.figs2{width:916px; margin:0 auto; overflow:hidden;}<br />
.fignew{font-size:13px; width:418px; margin:10px auto; float:left; padding:0 20px 0 20px;}<br />
.fig{font-size:13px; width:500px; margin:10px auto;}<br />
</style><br />
<br><br />
<br />
<div style="text-align:center;font-size:34px;color:white;"><b>Light-Activated Cell Lysis </b></div> <br />
<br><br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Objectives</div></b><br />
<div style="padding-right:150px;"><img src="https://static.igem.org/mediawiki/2012/f/fc/Lightdispschematic.gif" height="500" width="700" /></div><br />
<br><br />
<p style="color:black;text-indent:30px;">In order to develop a module for light activated cell lysis, we had to implement two elements:<br />
<ol style="font-size:15px"><li><b>Construct a light-activation system that can express a downstream gene of interest.</b></li><br />
<li><b>Express a cytolytic protein that can be expressed as our therapeutic drug to lyse cancer cells.</b></li><br />
</ul><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 1: Light-Activated Sensor</div></b><br><br><br />
<b><div class="name" align="center" style="font-size:16px;">Selection of YF1/FixJ Blue Light Sensor</div></b><br><br />
<p style="color:black;text-indent:30px;">After reading many papers to select an appropriate light-sensing system to use, we selected the YF1/FixJ blue light system. We had also considered the red light sensor Cph8 but ultimately decided on YF1/FixJ because of its high on/off ratio of gene expression and also because of its availability to us (we were fortunate enough to come across the YF1/FixJ system in the form of the pDawn plasmid from the Moglich lab in Germany).</p><br><br />
<b><div class="name" align="center" style="font-size:16px;">YF1/FixJ System (pDawn)</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">As shown below in Figure 1, the YF1/FixJ system works through a "repress the repressor" concept. Upon 480 nm blue light illumination, YF1 (a fusion of a LOV protein domain and a histidine kinase) phosphorylates a FixJ response regulator that activates the pFixK2 promoter. The activation of pFixK2, promotes expression of the cI repressor that, in turn, represses the lambda promoter pR. The net result is activation of the gene in the downstream MCS. </p><br><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/7/74/PDAWN.gif" /><br />
</div><br />
<br><br />
<div style="text-align:center"><b>Figure 1</b><br /></div><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 2: Expression of a Cytolytic Protein</div></b><br><br />
<b><div class="name" align="center">Cytolysin A (ClyA)</div></b><br><br />
<p style="color:black;text-indent:30px;"><br />
ClyA is a protein native to E. coli, Shigella flexneri, and Salmonella typhi that is capable of forming 13-mer pore complexes in a redox-independent manner. Expression of clyA in the absence of other hemolytic toxins is sufficient to induce hemolysis experimentally, and is therefore considered to be a potent cytolytic agent. Unlike a similar protein, HlyA, ClyA is not synthesized as a protoxin, which requires further posttranslational modifications to become active. ClyA is functional immediately following translation of mRNA to protein.<br />
<br />
ClyA is a 34kDa protein that is composed primarily of α-helical bundles that form a rod-shaped molecule. The membrane insertion domain is known as a β tongue (shown in yellow in Figure 2) and is critical for hemolytic activity. If the β tongue is mutated, the hemolytic activity of clyA is abrogated. </p><br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/2/22/ClyAPoregif.gif" /><br><br />
<br><b>Figure 2</b></div>Figure 2: ClyA forms a 13-mer pore complex that consists of hydrophobic beta tongues (yellow) on the head domains of individual monomer units that play an important role in influencing its cytolytic functions.<br />
</div><br />
<br />
<br><br />
<b><div class="name" align="center">Mechanism of Action</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">We selected ClyA as our cytolytic protein because of its unique mechanism of action that makes it especially potent. ClyA is secreted from bacteria in outer membrane vesicles (OMV's), within which it forms pore assemblies. As shown below in Figure 3, these pore assemblies allow ClyA to latch on to the cell wall of other cells and through the use of its encapsulating pore assembly lyse the cell wall. </p><br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/6/6c/ClyA-Pore-Assembly.jpg" /><br><br />
<br><b>Figure 3</b></div>Figure 3: Shown above in the first image are pore assemblies containing 13-mers of ClyA interacting with the surface of the target membrane. The second image below shows the ClyA assembly lysing the cell membrane through pore formation. (Wallace et. al 2000)<br />
</div><br />
<br />
<br />
<br />
</body><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/LightActivatedOverviewTeam:Penn/LightActivatedOverview2012-10-27T03:31:13Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
.figs2{width:916px; margin:0 auto; overflow:hidden;}<br />
.fignew{font-size:13px; width:418px; margin:10px auto; float:left; padding:0 20px 0 20px;}<br />
.fig{font-size:13px; width:500px; margin:10px auto;}<br />
</style><br />
<br><br />
<br />
<div style="text-align:center;font-size:34px;color:white;"><b>Light-Activated Cell Lysis </b></div> <br />
<br><br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Objectives</div></b><br />
<div class="fig"><div align="center" style="padding-right:150px;"><img src="https://static.igem.org/mediawiki/2012/f/fc/Lightdispschematic.gif" height="500" width="700" /></div></div><br />
<br><br />
<p style="color:black;text-indent:30px;">In order to develop a module for light activated cell lysis, we had to implement two elements:<br />
<ol style="font-size:15px"><li><b>Construct a light-activation system that can express a downstream gene of interest.</b></li><br />
<li><b>Express a cytolytic protein that can be expressed as our therapeutic drug to lyse cancer cells.</b></li><br />
</ul><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 1: Light-Activated Sensor</div></b><br><br><br />
<b><div class="name" align="center" style="font-size:16px;">Selection of YF1/FixJ Blue Light Sensor</div></b><br><br />
<p style="color:black;text-indent:30px;">After reading many papers to select an appropriate light-sensing system to use, we selected the YF1/FixJ blue light system. We had also considered the red light sensor Cph8 but ultimately decided on YF1/FixJ because of its high on/off ratio of gene expression and also because of its availability to us (we were fortunate enough to come across the YF1/FixJ system in the form of the pDawn plasmid from the Moglich lab in Germany).</p><br><br />
<b><div class="name" align="center" style="font-size:16px;">YF1/FixJ System (pDawn)</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">As shown below in Figure 1, the YF1/FixJ system works through a "repress the repressor" concept. Upon 480 nm blue light illumination, YF1 (a fusion of a LOV protein domain and a histidine kinase) phosphorylates a FixJ response regulator that activates the pFixK2 promoter. The activation of pFixK2, promotes expression of the cI repressor that, in turn, represses the lambda promoter pR. The net result is activation of the gene in the downstream MCS. </p><br><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/7/74/PDAWN.gif" /><br />
</div><br />
<br><br />
<div style="text-align:center"><b>Figure 1</b><br /></div><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 2: Expression of a Cytolytic Protein</div></b><br><br />
<b><div class="name" align="center">Cytolysin A (ClyA)</div></b><br><br />
<p style="color:black;text-indent:30px;"><br />
ClyA is a protein native to E. coli, Shigella flexneri, and Salmonella typhi that is capable of forming 13-mer pore complexes in a redox-independent manner. Expression of clyA in the absence of other hemolytic toxins is sufficient to induce hemolysis experimentally, and is therefore considered to be a potent cytolytic agent. Unlike a similar protein, HlyA, ClyA is not synthesized as a protoxin, which requires further posttranslational modifications to become active. ClyA is functional immediately following translation of mRNA to protein.<br />
<br />
ClyA is a 34kDa protein that is composed primarily of α-helical bundles that form a rod-shaped molecule. The membrane insertion domain is known as a β tongue (shown in yellow in Figure 2) and is critical for hemolytic activity. If the β tongue is mutated, the hemolytic activity of clyA is abrogated. </p><br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/2/22/ClyAPoregif.gif" /><br><br />
<br><b>Figure 2</b></div>Figure 2: ClyA forms a 13-mer pore complex that consists of hydrophobic beta tongues (yellow) on the head domains of individual monomer units that play an important role in influencing its cytolytic functions.<br />
</div><br />
<br />
<br><br />
<b><div class="name" align="center">Mechanism of Action</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">We selected ClyA as our cytolytic protein because of its unique mechanism of action that makes it especially potent. ClyA is secreted from bacteria in outer membrane vesicles (OMV's), within which it forms pore assemblies. As shown below in Figure 3, these pore assemblies allow ClyA to latch on to the cell wall of other cells and through the use of its encapsulating pore assembly lyse the cell wall. </p><br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/6/6c/ClyA-Pore-Assembly.jpg" /><br><br />
<br><b>Figure 3</b></div>Figure 3: Shown above in the first image are pore assemblies containing 13-mers of ClyA interacting with the surface of the target membrane. The second image below shows the ClyA assembly lysing the cell membrane through pore formation. (Wallace et. al 2000)<br />
</div><br />
<br />
<br />
<br />
</body><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/LightActivatedOverviewTeam:Penn/LightActivatedOverview2012-10-27T03:31:03Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
.figs2{width:916px; margin:0 auto; overflow:hidden;}<br />
.fignew{font-size:13px; width:418px; margin:10px auto; float:left; padding:0 20px 0 20px;}<br />
.fig{font-size:13px; width:500px; margin:10px auto;}<br />
</style><br />
<br><br />
<br />
<div style="text-align:center;font-size:34px;color:white;"><b>Light-Activated Cell Lysis </b></div> <br />
<br><br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Objectives</div></b><br />
<div class="fig"><div align="center" style="padding-right:100px;"><img src="https://static.igem.org/mediawiki/2012/f/fc/Lightdispschematic.gif" height="500" width="700" /></div></div><br />
<br><br />
<p style="color:black;text-indent:30px;">In order to develop a module for light activated cell lysis, we had to implement two elements:<br />
<ol style="font-size:15px"><li><b>Construct a light-activation system that can express a downstream gene of interest.</b></li><br />
<li><b>Express a cytolytic protein that can be expressed as our therapeutic drug to lyse cancer cells.</b></li><br />
</ul><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 1: Light-Activated Sensor</div></b><br><br><br />
<b><div class="name" align="center" style="font-size:16px;">Selection of YF1/FixJ Blue Light Sensor</div></b><br><br />
<p style="color:black;text-indent:30px;">After reading many papers to select an appropriate light-sensing system to use, we selected the YF1/FixJ blue light system. We had also considered the red light sensor Cph8 but ultimately decided on YF1/FixJ because of its high on/off ratio of gene expression and also because of its availability to us (we were fortunate enough to come across the YF1/FixJ system in the form of the pDawn plasmid from the Moglich lab in Germany).</p><br><br />
<b><div class="name" align="center" style="font-size:16px;">YF1/FixJ System (pDawn)</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">As shown below in Figure 1, the YF1/FixJ system works through a "repress the repressor" concept. Upon 480 nm blue light illumination, YF1 (a fusion of a LOV protein domain and a histidine kinase) phosphorylates a FixJ response regulator that activates the pFixK2 promoter. The activation of pFixK2, promotes expression of the cI repressor that, in turn, represses the lambda promoter pR. The net result is activation of the gene in the downstream MCS. </p><br><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/7/74/PDAWN.gif" /><br />
</div><br />
<br><br />
<div style="text-align:center"><b>Figure 1</b><br /></div><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 2: Expression of a Cytolytic Protein</div></b><br><br />
<b><div class="name" align="center">Cytolysin A (ClyA)</div></b><br><br />
<p style="color:black;text-indent:30px;"><br />
ClyA is a protein native to E. coli, Shigella flexneri, and Salmonella typhi that is capable of forming 13-mer pore complexes in a redox-independent manner. Expression of clyA in the absence of other hemolytic toxins is sufficient to induce hemolysis experimentally, and is therefore considered to be a potent cytolytic agent. Unlike a similar protein, HlyA, ClyA is not synthesized as a protoxin, which requires further posttranslational modifications to become active. ClyA is functional immediately following translation of mRNA to protein.<br />
<br />
ClyA is a 34kDa protein that is composed primarily of α-helical bundles that form a rod-shaped molecule. The membrane insertion domain is known as a β tongue (shown in yellow in Figure 2) and is critical for hemolytic activity. If the β tongue is mutated, the hemolytic activity of clyA is abrogated. </p><br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/2/22/ClyAPoregif.gif" /><br><br />
<br><b>Figure 2</b></div>Figure 2: ClyA forms a 13-mer pore complex that consists of hydrophobic beta tongues (yellow) on the head domains of individual monomer units that play an important role in influencing its cytolytic functions.<br />
</div><br />
<br />
<br><br />
<b><div class="name" align="center">Mechanism of Action</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">We selected ClyA as our cytolytic protein because of its unique mechanism of action that makes it especially potent. ClyA is secreted from bacteria in outer membrane vesicles (OMV's), within which it forms pore assemblies. As shown below in Figure 3, these pore assemblies allow ClyA to latch on to the cell wall of other cells and through the use of its encapsulating pore assembly lyse the cell wall. </p><br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/6/6c/ClyA-Pore-Assembly.jpg" /><br><br />
<br><b>Figure 3</b></div>Figure 3: Shown above in the first image are pore assemblies containing 13-mers of ClyA interacting with the surface of the target membrane. The second image below shows the ClyA assembly lysing the cell membrane through pore formation. (Wallace et. al 2000)<br />
</div><br />
<br />
<br />
<br />
</body><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/LightActivatedOverviewTeam:Penn/LightActivatedOverview2012-10-27T03:30:38Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
.figs2{width:916px; margin:0 auto; overflow:hidden;}<br />
.fignew{font-size:13px; width:418px; margin:10px auto; float:left; padding:0 20px 0 20px;}<br />
.fig{font-size:13px; width:500px; margin:10px auto;}<br />
</style><br />
<br><br />
<br />
<div style="text-align:center;font-size:34px;color:white;"><b>Light-Activated Cell Lysis </b></div> <br />
<br><br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Objectives</div></b><br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/f/fc/Lightdispschematic.gif" height="500" width="700" /></div></div><br />
<br><br />
<p style="color:black;text-indent:30px;">In order to develop a module for light activated cell lysis, we had to implement two elements:<br />
<ol style="font-size:15px"><li><b>Construct a light-activation system that can express a downstream gene of interest.</b></li><br />
<li><b>Express a cytolytic protein that can be expressed as our therapeutic drug to lyse cancer cells.</b></li><br />
</ul><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 1: Light-Activated Sensor</div></b><br><br><br />
<b><div class="name" align="center" style="font-size:16px;">Selection of YF1/FixJ Blue Light Sensor</div></b><br><br />
<p style="color:black;text-indent:30px;">After reading many papers to select an appropriate light-sensing system to use, we selected the YF1/FixJ blue light system. We had also considered the red light sensor Cph8 but ultimately decided on YF1/FixJ because of its high on/off ratio of gene expression and also because of its availability to us (we were fortunate enough to come across the YF1/FixJ system in the form of the pDawn plasmid from the Moglich lab in Germany).</p><br><br />
<b><div class="name" align="center" style="font-size:16px;">YF1/FixJ System (pDawn)</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">As shown below in Figure 1, the YF1/FixJ system works through a "repress the repressor" concept. Upon 480 nm blue light illumination, YF1 (a fusion of a LOV protein domain and a histidine kinase) phosphorylates a FixJ response regulator that activates the pFixK2 promoter. The activation of pFixK2, promotes expression of the cI repressor that, in turn, represses the lambda promoter pR. The net result is activation of the gene in the downstream MCS. </p><br><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/7/74/PDAWN.gif" /><br />
</div><br />
<br><br />
<div style="text-align:center"><b>Figure 1</b><br /></div><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 2: Expression of a Cytolytic Protein</div></b><br><br />
<b><div class="name" align="center">Cytolysin A (ClyA)</div></b><br><br />
<p style="color:black;text-indent:30px;"><br />
ClyA is a protein native to E. coli, Shigella flexneri, and Salmonella typhi that is capable of forming 13-mer pore complexes in a redox-independent manner. Expression of clyA in the absence of other hemolytic toxins is sufficient to induce hemolysis experimentally, and is therefore considered to be a potent cytolytic agent. Unlike a similar protein, HlyA, ClyA is not synthesized as a protoxin, which requires further posttranslational modifications to become active. ClyA is functional immediately following translation of mRNA to protein.<br />
<br />
ClyA is a 34kDa protein that is composed primarily of α-helical bundles that form a rod-shaped molecule. The membrane insertion domain is known as a β tongue (shown in yellow in Figure 2) and is critical for hemolytic activity. If the β tongue is mutated, the hemolytic activity of clyA is abrogated. </p><br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/2/22/ClyAPoregif.gif" /><br><br />
<br><b>Figure 2</b></div>Figure 2: ClyA forms a 13-mer pore complex that consists of hydrophobic beta tongues (yellow) on the head domains of individual monomer units that play an important role in influencing its cytolytic functions.<br />
</div><br />
<br />
<br><br />
<b><div class="name" align="center">Mechanism of Action</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">We selected ClyA as our cytolytic protein because of its unique mechanism of action that makes it especially potent. ClyA is secreted from bacteria in outer membrane vesicles (OMV's), within which it forms pore assemblies. As shown below in Figure 3, these pore assemblies allow ClyA to latch on to the cell wall of other cells and through the use of its encapsulating pore assembly lyse the cell wall. </p><br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/6/6c/ClyA-Pore-Assembly.jpg" /><br><br />
<br><b>Figure 3</b></div>Figure 3: Shown above in the first image are pore assemblies containing 13-mers of ClyA interacting with the surface of the target membrane. The second image below shows the ClyA assembly lysing the cell membrane through pore formation. (Wallace et. al 2000)<br />
</div><br />
<br />
<br />
<br />
</body><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/LightActivatedOverviewTeam:Penn/LightActivatedOverview2012-10-27T03:30:20Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
.figs2{width:916px; margin:0 auto; overflow:hidden;}<br />
.fignew{font-size:13px; width:418px; margin:10px auto; float:left; padding:0 20px 0 20px;}<br />
.fig{font-size:13px; width:500px; margin:10px auto;}<br />
</style><br />
<br><br />
<br />
<div style="text-align:center;font-size:34px;color:white;"><b>Light-Activated Cell Lysis </b></div> <br />
<br><br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Objectives</div></b><br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/f/fc/Lightdispschematic.gif" height="300" width="500" /></div></div><br />
<br><br />
<p style="color:black;text-indent:30px;">In order to develop a module for light activated cell lysis, we had to implement two elements:<br />
<ol style="font-size:15px"><li><b>Construct a light-activation system that can express a downstream gene of interest.</b></li><br />
<li><b>Express a cytolytic protein that can be expressed as our therapeutic drug to lyse cancer cells.</b></li><br />
</ul><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 1: Light-Activated Sensor</div></b><br><br><br />
<b><div class="name" align="center" style="font-size:16px;">Selection of YF1/FixJ Blue Light Sensor</div></b><br><br />
<p style="color:black;text-indent:30px;">After reading many papers to select an appropriate light-sensing system to use, we selected the YF1/FixJ blue light system. We had also considered the red light sensor Cph8 but ultimately decided on YF1/FixJ because of its high on/off ratio of gene expression and also because of its availability to us (we were fortunate enough to come across the YF1/FixJ system in the form of the pDawn plasmid from the Moglich lab in Germany).</p><br><br />
<b><div class="name" align="center" style="font-size:16px;">YF1/FixJ System (pDawn)</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">As shown below in Figure 1, the YF1/FixJ system works through a "repress the repressor" concept. Upon 480 nm blue light illumination, YF1 (a fusion of a LOV protein domain and a histidine kinase) phosphorylates a FixJ response regulator that activates the pFixK2 promoter. The activation of pFixK2, promotes expression of the cI repressor that, in turn, represses the lambda promoter pR. The net result is activation of the gene in the downstream MCS. </p><br><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/7/74/PDAWN.gif" /><br />
</div><br />
<br><br />
<div style="text-align:center"><b>Figure 1</b><br /></div><br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objective 2: Expression of a Cytolytic Protein</div></b><br><br />
<b><div class="name" align="center">Cytolysin A (ClyA)</div></b><br><br />
<p style="color:black;text-indent:30px;"><br />
ClyA is a protein native to E. coli, Shigella flexneri, and Salmonella typhi that is capable of forming 13-mer pore complexes in a redox-independent manner. Expression of clyA in the absence of other hemolytic toxins is sufficient to induce hemolysis experimentally, and is therefore considered to be a potent cytolytic agent. Unlike a similar protein, HlyA, ClyA is not synthesized as a protoxin, which requires further posttranslational modifications to become active. ClyA is functional immediately following translation of mRNA to protein.<br />
<br />
ClyA is a 34kDa protein that is composed primarily of α-helical bundles that form a rod-shaped molecule. The membrane insertion domain is known as a β tongue (shown in yellow in Figure 2) and is critical for hemolytic activity. If the β tongue is mutated, the hemolytic activity of clyA is abrogated. </p><br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/2/22/ClyAPoregif.gif" /><br><br />
<br><b>Figure 2</b></div>Figure 2: ClyA forms a 13-mer pore complex that consists of hydrophobic beta tongues (yellow) on the head domains of individual monomer units that play an important role in influencing its cytolytic functions.<br />
</div><br />
<br />
<br><br />
<b><div class="name" align="center">Mechanism of Action</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">We selected ClyA as our cytolytic protein because of its unique mechanism of action that makes it especially potent. ClyA is secreted from bacteria in outer membrane vesicles (OMV's), within which it forms pore assemblies. As shown below in Figure 3, these pore assemblies allow ClyA to latch on to the cell wall of other cells and through the use of its encapsulating pore assembly lyse the cell wall. </p><br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/6/6c/ClyA-Pore-Assembly.jpg" /><br><br />
<br><b>Figure 3</b></div>Figure 3: Shown above in the first image are pore assemblies containing 13-mers of ClyA interacting with the surface of the target membrane. The second image below shows the ClyA assembly lysing the cell membrane through pore formation. (Wallace et. al 2000)<br />
</div><br />
<br />
<br />
<br />
</body><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/SurfaceDisplayOverviewTeam:Penn/SurfaceDisplayOverview2012-10-27T03:28:52Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
.figs2{width:916px; margin:0 auto; overflow:hidden;}<br />
.fignew{font-size:13px; width:418px; margin:10px auto; float:left; padding:0 20px 0 20px;}<br />
.fig{font-size:13px; width:500px; margin:10px auto;}<br />
</style><br />
<br><br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Surface Display and Targeting</div></b><br><br />
<br />
<div class="fig"><div align="center"><img style="align:center;"src="https://static.igem.org/mediawiki/2012/a/a4/Surfacedispschematic.gif" height="300" width="500"></div></div><br />
While engineering bacteria as a light-activated drug delivery <a href="https://2012.igem.org/Team:Penn/LightActivatedOverview "> system </a> is novel and useful by itself, the targeting potential of our light-activated bacterial therapeutic would be greatly improved if the bacteria also could target specific cells. The goal of the second module of our system is to allow bacterial targeting to cancer cells. We sought to achieve this by displaying an engineered cancer cell binding protein on the surface of our <i> E. coli</i>. We successfully displayed DARPin H10-2-G3, an antibody-mimetic protein rationally evolved to picomolar affinity with HER2, a breast cancer biomarker. We verified that bacteria displaying this protein can selectively bind to breast cancer cells <i> in vitro</i>. This class of protein has not been displayed before on the surface of bacteria. Along the way, we developed a generalized BioBrick surface display system which allows future iGEM teams to display proteins of their choosing on the surface of bacteria.<br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objectives</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">Our goals were twofold:<br />
<ol style="font-size:15px"><li><b> Achieve the first display of DARPin H10-2-G3 on the surface of <i> E. coli</i> and verify that the system targets cancer cells. </b></li><br />
<li><b> Create a generalized BioBrick surface display platform for other labs and iGEM teams.</b></li><br />
</ul><br />
</div><br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Ice Nucleation Protein</div></b><br />
<br><br />
We chose to use the Ice Nucleation Protein (Figure 1) as our surface display carrier protein. The Ice Nucleation Protein protein has been used to display enzymes [1], typically for biocatalysis applications (e.g. Edinburgh iGEM 2011). To make the protein a more manageable size, we truncated the protein to the N and C terminal domains only. The C terminal domain is displayed at the cell surface, while the N terminal domain remains in the outer membrane. We sought to apply this system to health/medicine by displaying DARPin H10-2-G3.<br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/e/e4/INPNC.png" /><br><br />
<b>Figure 1</b></div><div style="text-align:center">Figure 1: Proposed Structure of Ice Nucleation Protein.</div></div><br />
<p style="color:black"><br><br><br />
[1] Zahnd, C., Wyler, E., Schwenk, J. M., Steiner, D., Lawrence, M. C., McKern, N. M., Pecorari, F., et al. (2007). A designed ankyrin repeat protein evolved to picomolar affinity to Her2. Journal of molecular biology, 369(4), 1015–28. doi:10.1016/j.jmb.2007.03.028 <br />
</p><br />
</div><br />
<br><br />
<br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">DARPin H10-2-G3</div></b><br />
<br><br />
The <a href="http://www.bioc.uzh.ch/plueckthun/"> lab </a> of Andreas Plueckthun at ETH Zurich has pioneered the development of Designed Ankyrin Repeat Protein (DARPin) technology. DARPins are engineered antibody-mimetic proteins consisting of 3-5 ankyrin repeat motifs. These proteins have been rationally evolved to high binding affinity with targets through ribosome display. In 2007, the group developed H10-2-G3 [2], a 14.7kDa DARPin evolved to 90pM affinity with the extracellular domain of HER2 (Figure 2).<br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/b/b7/DARPingif.gif" /><br><br />
<b>Figure 2</b></div><div style="text-align:center">Figure 2: Structure of DARPin-H10-2-G3.</div></div><br />
<p style="color:black"><br><br><br />
[2] Zahnd, C., Wyler, E., Schwenk, J. M., Steiner, D., Lawrence, M. C., McKern, N. M., Pecorari, F., et al. (2007). A designed ankyrin repeat protein evolved to picomolar affinity to Her2. Journal of molecular biology, 369(4), 1015–28. doi:10.1016/j.jmb.2007.03.028 <br />
</p><br />
</div><br />
<br><br />
<br />
<br />
<br />
<br />
<br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/SurfaceDisplayOverviewTeam:Penn/SurfaceDisplayOverview2012-10-27T03:28:25Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:936px; padding:30px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
.figs2{width:916px; margin:0 auto; overflow:hidden;}<br />
.fignew{font-size:13px; width:418px; margin:10px auto; float:left; padding:0 20px 0 20px;}<br />
.fig{font-size:13px; width:500px; margin:10px auto;}<br />
</style><br />
<br><br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Surface Display and Targeting</div></b><br><br />
<br />
<div class="fig"><div align="center"><img style="align:center;"src="https://static.igem.org/mediawiki/2012/a/a4/Surfacedispschematic.gif"></div></div><br />
While engineering bacteria as a light-activated drug delivery <a href="https://2012.igem.org/Team:Penn/LightActivatedOverview "> system </a> is novel and useful by itself, the targeting potential of our light-activated bacterial therapeutic would be greatly improved if the bacteria also could target specific cells. The goal of the second module of our system is to allow bacterial targeting to cancer cells. We sought to achieve this by displaying an engineered cancer cell binding protein on the surface of our <i> E. coli</i>. We successfully displayed DARPin H10-2-G3, an antibody-mimetic protein rationally evolved to picomolar affinity with HER2, a breast cancer biomarker. We verified that bacteria displaying this protein can selectively bind to breast cancer cells <i> in vitro</i>. This class of protein has not been displayed before on the surface of bacteria. Along the way, we developed a generalized BioBrick surface display system which allows future iGEM teams to display proteins of their choosing on the surface of bacteria.<br />
</div><br />
<div class="bigbox"><br />
<b><div class="name" align="center">Objectives</div></b><br />
<br><br />
<p style="color:black;text-indent:30px;">Our goals were twofold:<br />
<ol style="font-size:15px"><li><b> Achieve the first display of DARPin H10-2-G3 on the surface of <i> E. coli</i> and verify that the system targets cancer cells. </b></li><br />
<li><b> Create a generalized BioBrick surface display platform for other labs and iGEM teams.</b></li><br />
</ul><br />
</div><br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">Ice Nucleation Protein</div></b><br />
<br><br />
We chose to use the Ice Nucleation Protein (Figure 1) as our surface display carrier protein. The Ice Nucleation Protein protein has been used to display enzymes [1], typically for biocatalysis applications (e.g. Edinburgh iGEM 2011). To make the protein a more manageable size, we truncated the protein to the N and C terminal domains only. The C terminal domain is displayed at the cell surface, while the N terminal domain remains in the outer membrane. We sought to apply this system to health/medicine by displaying DARPin H10-2-G3.<br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/e/e4/INPNC.png" /><br><br />
<b>Figure 1</b></div><div style="text-align:center">Figure 1: Proposed Structure of Ice Nucleation Protein.</div></div><br />
<p style="color:black"><br><br><br />
[1] Zahnd, C., Wyler, E., Schwenk, J. M., Steiner, D., Lawrence, M. C., McKern, N. M., Pecorari, F., et al. (2007). A designed ankyrin repeat protein evolved to picomolar affinity to Her2. Journal of molecular biology, 369(4), 1015–28. doi:10.1016/j.jmb.2007.03.028 <br />
</p><br />
</div><br />
<br><br />
<br />
<br />
<br />
<div class="bigbox"><br />
<b><div class="name" align="center">DARPin H10-2-G3</div></b><br />
<br><br />
The <a href="http://www.bioc.uzh.ch/plueckthun/"> lab </a> of Andreas Plueckthun at ETH Zurich has pioneered the development of Designed Ankyrin Repeat Protein (DARPin) technology. DARPins are engineered antibody-mimetic proteins consisting of 3-5 ankyrin repeat motifs. These proteins have been rationally evolved to high binding affinity with targets through ribosome display. In 2007, the group developed H10-2-G3 [2], a 14.7kDa DARPin evolved to 90pM affinity with the extracellular domain of HER2 (Figure 2).<br />
<br><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/b/b7/DARPingif.gif" /><br><br />
<b>Figure 2</b></div><div style="text-align:center">Figure 2: Structure of DARPin-H10-2-G3.</div></div><br />
<p style="color:black"><br><br><br />
[2] Zahnd, C., Wyler, E., Schwenk, J. M., Steiner, D., Lawrence, M. C., McKern, N. M., Pecorari, F., et al. (2007). A designed ankyrin repeat protein evolved to picomolar affinity to Her2. Journal of molecular biology, 369(4), 1015–28. doi:10.1016/j.jmb.2007.03.028 <br />
</p><br />
</div><br />
<br><br />
<br />
<br />
<br />
<br />
<br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/SurfaceDisplayBBaTeam:Penn/SurfaceDisplayBBa2012-10-27T03:26:49Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site}}<br />
<html><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #ffffff;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.pic1{ float:left; margin:0 40px 0 0; width:150px;}<br />
.name{ font-size:20px;}<br />
.figs2{width:916px; margin:0 auto; overflow:hidden;}<br />
.fignew{font-size:13px; width:418px; margin:10px auto; float:left; padding:0 20px 0 20px;}<br />
.fig{font-size:13px; width:500px; margin:10px auto;}<br />
</style><br />
<br><br />
<br />
<br />
<br />
<br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Display Your Own Proteins: BBa_K811005<br />
</div></b><br /><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/0/06/Your-Protein-Here.gif" width="250" height="350"/></div></div><br />
<br />
<br />
<br />
<p style="color:black;text-indent:30px;"><br />
Surface display has wide applications in industry and medicine. We wanted to create a simple system for any iGEM team or lab to display proteins on the surface of bacteria. Using INPNC and our 12aa GS linker, we created INPNC-MCS, a BioBrick which allows teams to clone in any INPNC fusion partner of their choosing and have it displayed on the surface of bacteria. This system only requires a one step ligation with BamHI and PstI cloning sites. <b>This <a href="http://partsregistry.org/Part:BBa_K811005">BioBrick</a> was recently awarded "Best BioBrick: Engineered" at the 2012 iGEM Americas East Regional Jamboree!</b> </p><br />
</div><br />
<br />
<br />
<br />
<div class="bigbox"><br />
<br />
<b><div class="name" align="center">Experience<br />
</div></b><br /><br />
<p style="color:black;text-indent:30px;"><br />
Part BBa_K811005 has been utilized by the 2012 Penn iGEM team to display a two completely different classes of proteins and we are currently using it to display a variety of other proteins. The red fluorescent protein mCherry has been successfully displayed on the surface of E. Coli, where it can produce fluorescence. We displayed mCherry on the outer membrane of E. coli BL21. After sonication and centrifugation of induced cells, almost all of the mCherry was localized in the membrane fraction when fused to INPNC, whereas in the control Intein-mCherry fusion (which exhibits cytoplasmic localization), all of the mCherry was contained in the lysate (Figure 1).</p><br />
<br />
<div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/a/a2/MCherry-vs-INPNC-mCherry.jpg" width="250" height="350"/><br><br />
<b>Figure 1</b></div>Figure 1: Surface display of mCherry using INPNC system. INPNC-mCherry and Intein-mCherry fusions were expressed in E. coli BL21 in the pET26b expression vector and Wood-Intein expression plasmid, respectively. When fused to INPNC, almost all mCherry was localized in the membrane fraction after sonication and centrifugation, while in the case of Intein-mCherry, all mCherry was localized in the cytoplasmic lysate.</div><br />
<br />
<br />
Furthermore, a HER2 binding protein, DARPin H20-2-G3 has also displayed on the surface of E. Coli, and has been shown to retain its HER2 binding affinity upon surface display through Part BBa_K811005.<br />
</div><br />
<br />
<br />
</html></div>Amurthurhttp://2012.igem.org/File:Surfdispmodule.gifFile:Surfdispmodule.gif2012-10-27T03:25:34Z<p>Amurthur: </p>
<hr />
<div></div>Amurthurhttp://2012.igem.org/File:Lightactivationmodule.gifFile:Lightactivationmodule.gif2012-10-27T03:24:56Z<p>Amurthur: </p>
<hr />
<div></div>Amurthurhttp://2012.igem.org/Team:Penn/VerifiGEMTeam:Penn/VerifiGEM2012-10-27T03:22:17Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<style type="text/css"><br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<div class="bigbox"><br />
<b><div class="name" align="center">The Question: How Can We Increase the Accuracy of Biobrick Parts Submitted to the Registry?</div></b><br />
<br><br><br />
<b><div class="name" align="center">Our Solution: VerifiGEM</div></b><br />
</div><br />
<div style="padding-left:150px;"><br />
<div class="prezi-player"><style type="text/css" media="screen">.prezi-player { width: 700px; } .prezi-player-links { text-align: center; }</style><object id="prezi_xlruzy2n7spo" name="prezi_xlruzy2n7spo" classid="clsid:D27CDB6E-AE6D-11cf-96B8-444553540000" width="700" height="500"><param name="movie" value="http://prezi.com/bin/preziloader.swf"/><param name="allowfullscreen" value="true"/><param name="allowFullScreenInteractive" value="true"/><param name="allowscriptaccess" value="always"/><param name="wmode" value="direct"/><param name="bgcolor" value="#ffffff"/><param name="flashvars" value="prezi_id=xlruzy2n7spo&amp;lock_to_path=0&amp;color=ffffff&amp;autoplay=no&amp;autohide_ctrls=0"/><embed id="preziEmbed_xlruzy2n7spo" name="preziEmbed_xlruzy2n7spo" src="http://prezi.com/bin/preziloader.swf" type="application/x-shockwave-flash" allowfullscreen="true" allowFullScreenInteractive="true" allowscriptaccess="always" width="700" height="500" bgcolor="#ffffff" flashvars="prezi_id=xlruzy2n7spo&amp;lock_to_path=0&amp;color=ffffff&amp;autoplay=no&amp;autohide_ctrls=0"></embed></object><div class="prezi-player-links"><p><a title="VerifiGEM " href="http://prezi.com/xlruzy2n7spo/verifigem/">VerifiGEM </a> on <a href="http://prezi.com">Prezi</a></p></div></div></div><br />
<br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/VerifiGEMTeam:Penn/VerifiGEM2012-10-27T03:21:44Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<style type="text/css"><br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<div class="bigbox"><br />
<b><div class="name" align="center">The Question: How Can We Increase the Accuracy of Biobrick Parts Submitted to the Registry?</div></b><br />
<br><br><br />
<b><div class="name" align="center">Our Solution: VerifiGEM</div></b><br />
</div><br />
<div style="padding-left:225px;"><br />
<div class="prezi-player"><style type="text/css" media="screen">.prezi-player { width: 550px; } .prezi-player-links { text-align: center; }</style><object id="prezi_xlruzy2n7spo" name="prezi_xlruzy2n7spo" classid="clsid:D27CDB6E-AE6D-11cf-96B8-444553540000" width="700" height="500"><param name="movie" value="http://prezi.com/bin/preziloader.swf"/><param name="allowfullscreen" value="true"/><param name="allowFullScreenInteractive" value="true"/><param name="allowscriptaccess" value="always"/><param name="wmode" value="direct"/><param name="bgcolor" value="#ffffff"/><param name="flashvars" value="prezi_id=xlruzy2n7spo&amp;lock_to_path=0&amp;color=ffffff&amp;autoplay=no&amp;autohide_ctrls=0"/><embed id="preziEmbed_xlruzy2n7spo" name="preziEmbed_xlruzy2n7spo" src="http://prezi.com/bin/preziloader.swf" type="application/x-shockwave-flash" allowfullscreen="true" allowFullScreenInteractive="true" allowscriptaccess="always" width="550" height="400" bgcolor="#ffffff" flashvars="prezi_id=xlruzy2n7spo&amp;lock_to_path=0&amp;color=ffffff&amp;autoplay=no&amp;autohide_ctrls=0"></embed></object><div class="prezi-player-links"><p><a title="VerifiGEM " href="http://prezi.com/xlruzy2n7spo/verifigem/"></a><a href="http://prezi.com"></a></p></div></div></div><br />
<br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/VerifiGEMTeam:Penn/VerifiGEM2012-10-27T03:21:23Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<style type="text/css"><br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<div class="bigbox"><br />
<b><div class="name" align="center">The Question: How Can We Increase the Accuracy of Biobrick Parts Submitted to the Registry?</div></b><br />
<br><br><br />
<b><div class="name" align="center">Our Solution: VerifiGEM</div></b><br />
</div><br />
<div style="padding-left:225px;"><br />
<div class="prezi-player"><style type="text/css" media="screen">.prezi-player { width: 550px; } .prezi-player-links { text-align: center; }</style><object id="prezi_xlruzy2n7spo" name="prezi_xlruzy2n7spo" classid="clsid:D27CDB6E-AE6D-11cf-96B8-444553540000" width="550" height="400"><param name="movie" value="http://prezi.com/bin/preziloader.swf"/><param name="allowfullscreen" value="true"/><param name="allowFullScreenInteractive" value="true"/><param name="allowscriptaccess" value="always"/><param name="wmode" value="direct"/><param name="bgcolor" value="#ffffff"/><param name="flashvars" value="prezi_id=xlruzy2n7spo&amp;lock_to_path=0&amp;color=ffffff&amp;autoplay=no&amp;autohide_ctrls=0"/><embed id="preziEmbed_xlruzy2n7spo" name="preziEmbed_xlruzy2n7spo" src="http://prezi.com/bin/preziloader.swf" type="application/x-shockwave-flash" allowfullscreen="true" allowFullScreenInteractive="true" allowscriptaccess="always" width="550" height="400" bgcolor="#ffffff" flashvars="prezi_id=xlruzy2n7spo&amp;lock_to_path=0&amp;color=ffffff&amp;autoplay=no&amp;autohide_ctrls=0"></embed></object><div class="prezi-player-links"><p><a title="VerifiGEM " href="http://prezi.com/xlruzy2n7spo/verifigem/"></a><a href="http://prezi.com"></a></p></div></div></div><br />
<br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/VerifiGEMTeam:Penn/VerifiGEM2012-10-27T03:21:00Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<style type="text/css"><br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<div class="bigbox"><br />
<b><div class="name" align="center">The Question: How Can We Increase the Accuracy of Biobrick Parts Submitted to the Registry?</div></b><br />
<br><br><br />
<b><div class="name" align="center">Our Solution: VerifiGEM</div></b><br />
</div><br />
<div style="padding-left:225px;"><br />
<div class="prezi-player"><style type="text/css" media="screen">.prezi-player { width: 550px; } .prezi-player-links { text-align: center; }</style><object id="prezi_xlruzy2n7spo" name="prezi_xlruzy2n7spo" classid="clsid:D27CDB6E-AE6D-11cf-96B8-444553540000" width="550" height="400"><param name="movie" value="http://prezi.com/bin/preziloader.swf"/><param name="allowfullscreen" value="true"/><param name="allowFullScreenInteractive" value="true"/><param name="allowscriptaccess" value="always"/><param name="wmode" value="direct"/><param name="bgcolor" value="#ffffff"/><param name="flashvars" value="prezi_id=xlruzy2n7spo&amp;lock_to_path=0&amp;color=ffffff&amp;autoplay=no&amp;autohide_ctrls=0"/><embed id="preziEmbed_xlruzy2n7spo" name="preziEmbed_xlruzy2n7spo" src="http://prezi.com/bin/preziloader.swf" type="application/x-shockwave-flash" allowfullscreen="true" allowFullScreenInteractive="true" allowscriptaccess="always" width="550" height="400" bgcolor="#ffffff" flashvars="prezi_id=xlruzy2n7spo&amp;lock_to_path=0&amp;color=ffffff&amp;autoplay=no&amp;autohide_ctrls=0"></embed></object><div class="prezi-player-links"><p><a title="VerifiGEM " href="http://prezi.com/xlruzy2n7spo/verifigem/">VerifiGEM </a> on <a href="http://prezi.com">Prezi</a></p></div></div></div><br />
<br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/VerifiGEMTeam:Penn/VerifiGEM2012-10-27T03:20:35Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<style type="text/css"><br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify; margin:0 0 20px 0;}<br />
.name{ font-size:20px;}<br />
</style><br />
<br><br />
<div class="bigbox"><br />
<b><div class="name" align="center">The Question: How Can We Increase the Accuracy of Biobrick Parts Submitted to the Registry?</div></b><br />
<br><br><br />
<b><div class="name" align="center">Our Solution: VerifiGEM</div></b><br />
</div><br />
<br />
<div class="prezi-player"><style type="text/css" media="screen">.prezi-player { width: 550px; } .prezi-player-links { text-align: center; }</style><object id="prezi_xlruzy2n7spo" name="prezi_xlruzy2n7spo" classid="clsid:D27CDB6E-AE6D-11cf-96B8-444553540000" width="550" height="400"><param name="movie" value="http://prezi.com/bin/preziloader.swf"/><param name="allowfullscreen" value="true"/><param name="allowFullScreenInteractive" value="true"/><param name="allowscriptaccess" value="always"/><param name="wmode" value="direct"/><param name="bgcolor" value="#ffffff"/><param name="flashvars" value="prezi_id=xlruzy2n7spo&amp;lock_to_path=0&amp;color=ffffff&amp;autoplay=no&amp;autohide_ctrls=0"/><embed id="preziEmbed_xlruzy2n7spo" name="preziEmbed_xlruzy2n7spo" src="http://prezi.com/bin/preziloader.swf" type="application/x-shockwave-flash" allowfullscreen="true" allowFullScreenInteractive="true" allowscriptaccess="always" width="550" height="400" bgcolor="#ffffff" flashvars="prezi_id=xlruzy2n7spo&amp;lock_to_path=0&amp;color=ffffff&amp;autoplay=no&amp;autohide_ctrls=0"></embed></object><div class="prezi-player-links"><p><a title="VerifiGEM " href="http://prezi.com/xlruzy2n7spo/verifigem/">VerifiGEM </a> on <a href="http://prezi.com">Prezi</a></p></div></div><br />
<br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/TeamTeam:Penn/Team2012-10-27T03:19:15Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<!-----------------------------------------------------------------------------------------------------><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #000000;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.flpic{float:left; margin:0 0 0 0; width:150px;}<br />
.pic1{width:150px;}<br />
.name{ <br />
font-size:20px;<br />
}<br />
.flleft{ float:left; width:400px; margin:0 0 0 50px;}<br />
</style><br />
<br />
<br />
<div class="all"><br />
<br><br />
<div style="text-align:center;font-size:34px;color:white;"><b>Our Team</b></div><br><br />
<br />
<div class="bigbox"><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/7/7a/Penn-iGEM-2012-Team-Photo.jpg" height="500" width="700" /></div><br><br />
The Penn iGEM 2012 Team consists of 4 undergraduates, 3 advisors, and many others who have provided important contributions along the way. Together, we have learned a lot over the last few months. We have taught ourselves different protocols and developed new standards to synchronize our work. Through our participation in iGEM, we have collaborated to learn more about synthetic biology - from the initial days of cloning constructs to the final days of imaging and analysis. Our idea for spatio-temporal control of drug delivery first originated in late May after weeks of reading papers. Slowly we have been able to piece together different components of the system to help the project materialize to the system it is today. We believe our work in optogenetics and drug delivery has a promising future and we are excited to share our results at the World Championships! <br />
</div><br />
<br />
<br />
<br />
<div class="und">Undergraduates</div><br />
<br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="https://static.igem.org/mediawiki/2012/c/c6/Clark_Park_4.jpg" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Ashwin Amurthur</b><br />
<br><br />
<i>"Now we're rolling"</i><br></div><br />
<br><br />
Ashwin is a sophomore at the University of Pennsylvania studying bioengineering and management. He hopes to attend medical school in the future.<br />
<br><br><br />
<b>Natural lab habitat:</b> Running transformations<br><br />
<b>Best kept lab secret:</b> Never ran a PCR <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/9mbudvt">www.tinyurl.com/9mbudvt</a></b> <br />
</div></div><br />
<br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/283077_1448493822523_1538520160_31353808_6931705_n-e1317115984293-221x300.jpg" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Michael Magaraci</b><br />
<br><i>"Sorry guys, I overslept..."</i><br></div><br><br />
Mike is a senior at the University of Pennsylvania studying bioengineering and management. <br />
<br><br><br />
<b>Natural lab habitat:</b> At the PCR machine<br><br />
<b>Best kept lab secret:</b> <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/9rpe7ap">www.tinyurl.com/9rpe7ap</a></b> <br />
</div></div><br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/Untitled-e1317089819600.png" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Peter Qiao</b><br />
<br><i>"There's nothing mini about my miniprep yields"</i><br></div><br><br />
Peter is a junior at the University of Pennsylvania studying bioengineering. He hopes to pursue an M.D/Ph.D in the future.<br />
<br><br><br />
<b>Natural lab habitat:</b> In the hood passaging cells<Br><br />
<b>Best kept lab secret:</b> Triple ligation <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/8e2zj3l">www.tinyurl.com/8e2zj3l</a></b> <br />
<br />
</div></div><br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/P1000543-e1317095308771-221x300.jpg" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Avin Veerakumar</b><br />
<br><i>"Do you want to sleep or do you want to win?"</i><br></div><br><br />
Avin is a senior at the University of Pennsylvania studying Bioengineering and Management.<br />
<br><br><br />
<b>Natural lab habitat:</b> On the confocal<br><br />
<b>Best kept lab secret:</b> Never ran a PCR <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/9ahlsc4">www.tinyurl.com/9ahlsc4</a></b> <br />
</div></div><br />
<br />
<div class="und">Advisors</div><br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/sarkar_web-e1317090202691.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Dr. Casim A. Sarkar</b><br />
<br><br><br />
Dr. Casim A. Sarkar is the Principal Investigator of the Molecular Cell Engineering Laboratory at the University of Pennsylvania Department of Bioengineering. His research interests include molecular cell engineering, protein engineering, ligand/receptor binding and trafficking, cell signaling and decision making, and computational, synthetic, and systems biology. Dr. Sarkar received a PhD in Chemical Engineering with a minor in Computational Biology at MIT and a BS in Chemical Engineering at the University of Texas at Austin. <br />
</div></div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://www.med.upenn.edu/apps/my/images/faculty_pics/goul2044.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Dr. Mark Goulian</b><br />
<br><br><br />
Dr. Mark Goulian is the Edmund J. and Louise W. Kahn Endowed Term Professor of Biology at the University of Pennsylvania. His research is focused on the regulatory circuits that bacteria use to sense and respond to the environment. His other research interests include two-component signaling in E. coli and directed evolution of signaling circuits. Dr. Goulian received his PhD from Harvard University. </div><br />
</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/MillerJordan_Penn-Fellow-e1317190345626.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Dr. Jordan S. Miller</b><br />
<br><br><br />
Dr. Jordan S. Miller is a postdoctoral fellow at the University of Pennsylvania in Dr. Christopher S. Chen's Tissue Microfabrication Laboratory in the Department of Bioengineering. He is currently also a board member of Hive76 and before his time at Penn, he was a developer at RepRap and and an associate at PTV Sciences. Dr. Miller received a PhD from Rice University and earned his undergraduate degree from MIT.<br />
</div></div><br />
<div class="und">Acknowledgements</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/DaphneNg-e1317243791521-224x300.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Daphne Ng</b><br />
<br><br><br />
Daphne Ng is a PhD candidate at the University of Pennsylvania in Dr. Casim A. Sarkar's Molecular Cell Engineering Laboratory in the Department of Bioengineering. She graduated from Cornell University in 2008 with a B.S. in Chemical Engineering. The 2012 Penn iGEM team would like to thank Daphne for her contribution to the cloning and design process, as well as providing expression and cloning vectors.</div><br />
</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/najafshah.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Najaf A. Shah</b><br />
<br><br><br />
Najaf Shah is a PhD candidate at the University of Pennsylvania in Dr. Casim A. Sarkar's Molecular Cell Engineering Laboratory in the Department of Bioengineering. The 2012 Penn iGEM team would like to thank Najaf for providing input on cloning, input on system design, and access to advanced imaging facilities.</div><br />
</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/board-sevile.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Sevile Mannickarottu</b><br />
<br><br><br />
Sevile Mannickarottu is the Director of Bioengineering Instructional Laboratories at the University of Pennsylvania. Before his current post, he was the Philadelphia Operations Manager for Technology Education Awareness and a Project Electrical Engineer at Lutron Electronics. He received an MLA in Religious Studies and a BSc in Electric Engineering from the University of Pennsylvania and is currently pursuing a PhD in Religious Studies. The 2012 Penn iGEM team would like to thank Sevile for providing his facilities and equipment for use during the project.<br />
</div></div><br />
<div class="und">Contributions</div><br />
<br />
<div class="bigbox"><br />
The Penn iGEM team would also like to thank the following individuals for their significant contributions to the team this year:<br />
<ul style="font-size:17px;padding-left:20px; margin:0 0;"><br />
<li><b>Christopher Fang-Yen</b>, PhD, Assistant Professor of Bioengineering at University of Pennsylvania</li><br />
<li><b>Matthew Lazzara</b>, PhD, Assistant Professor of Bioengineering at University of Pennsylvania</li><br />
<li><b>Dan Cohen</b>, PhD, Postdoctoral Fellow at University of Pennsylvania</li><br />
<li><b>Henry Ma</b>, Engineer at University of Pennsylvania Bioengineering Instructional Laboratories</li><br />
<li><b>Karsticum Computing Inc.</b>, Software development company that was instrumental in the construction of VerifiGEM</li></ul><br />
</div><br />
</div><br />
</body><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/TeamTeam:Penn/Team2012-10-27T03:18:58Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<!-----------------------------------------------------------------------------------------------------><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #000000;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.flpic{float:left; margin:0 0 0 0; width:150px;}<br />
.pic1{width:150px;}<br />
.name{ <br />
font-size:20px;<br />
}<br />
.flleft{ float:left; width:400px; margin:0 0 0 50px;}<br />
</style><br />
<br />
<br />
<div class="all"><br />
<br><br />
<div style="text-align:center;font-size:34px;color:white;"><b>Our Team</b></div><br><br />
<br />
<div class="bigbox"><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/7/7a/Penn-iGEM-2012-Team-Photo.jpg" height="500" width="700" /></div><br><br />
The Penn iGEM 2012 Team consists of 4 undergraduates, 3 advisors, and many others who have provided important contributions along the way. Together, we have learned a lot over the last few months. We have taught ourselves different protocols and developed new standards to synchronize our work. Through our participation in iGEM, we have collaborated to learn more about synthetic biology - from the initial days of cloning constructs to the final days of imaging and analysis. Our idea for spatio-temporal control of drug delivery first originated in late May after weeks of reading papers. Slowly we have been able to piece together different components of the system to help the project materialize to the system it is today. We believe our work in optogenetics and drug delivery has a promising future and we are excited to share our results at the World Championships! <br />
</div><br />
<br />
<br />
<br />
<div class="und">Undergraduates</div><br />
<br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="https://static.igem.org/mediawiki/2012/c/c6/Clark_Park_4.jpg" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Ashwin Amurthur</b><br />
<br><br />
<i>"Now we're rolling"</i><br></div><br />
<br><br />
Ashwin is a sophomore at the University of Pennsylvania studying bioengineering and management. He hopes to attend medical school in the future.<br />
<br><br><br />
<b>Natural lab habitat:</b> Running transformationsbr><br />
<b>Best kept lab secret:</b> Never ran a PCR <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/9mbudvt">www.tinyurl.com/9mbudvt</a></b> <br />
</div></div><br />
<br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/283077_1448493822523_1538520160_31353808_6931705_n-e1317115984293-221x300.jpg" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Michael Magaraci</b><br />
<br><i>"Sorry guys, I overslept..."</i><br></div><br><br />
Mike is a senior at the University of Pennsylvania studying bioengineering and management. <br />
<br><br><br />
<b>Natural lab habitat:</b> At the PCR machine<br><br />
<b>Best kept lab secret:</b> <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/9rpe7ap">www.tinyurl.com/9rpe7ap</a></b> <br />
</div></div><br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/Untitled-e1317089819600.png" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Peter Qiao</b><br />
<br><i>"There's nothing mini about my miniprep yields"</i><br></div><br><br />
Peter is a junior at the University of Pennsylvania studying bioengineering. He hopes to pursue an M.D/Ph.D in the future.<br />
<br><br><br />
<b>Natural lab habitat:</b> In the hood passaging cells<Br><br />
<b>Best kept lab secret:</b> Triple ligation <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/8e2zj3l">www.tinyurl.com/8e2zj3l</a></b> <br />
<br />
</div></div><br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/P1000543-e1317095308771-221x300.jpg" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Avin Veerakumar</b><br />
<br><i>"Do you want to sleep or do you want to win?"</i><br></div><br><br />
Avin is a senior at the University of Pennsylvania studying Bioengineering and Management.<br />
<br><br><br />
<b>Natural lab habitat:</b> On the confocal<br><br />
<b>Best kept lab secret:</b> Never ran a PCR <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/9ahlsc4">www.tinyurl.com/9ahlsc4</a></b> <br />
</div></div><br />
<br />
<div class="und">Advisors</div><br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/sarkar_web-e1317090202691.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Dr. Casim A. Sarkar</b><br />
<br><br><br />
Dr. Casim A. Sarkar is the Principal Investigator of the Molecular Cell Engineering Laboratory at the University of Pennsylvania Department of Bioengineering. His research interests include molecular cell engineering, protein engineering, ligand/receptor binding and trafficking, cell signaling and decision making, and computational, synthetic, and systems biology. Dr. Sarkar received a PhD in Chemical Engineering with a minor in Computational Biology at MIT and a BS in Chemical Engineering at the University of Texas at Austin. <br />
</div></div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://www.med.upenn.edu/apps/my/images/faculty_pics/goul2044.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Dr. Mark Goulian</b><br />
<br><br><br />
Dr. Mark Goulian is the Edmund J. and Louise W. Kahn Endowed Term Professor of Biology at the University of Pennsylvania. His research is focused on the regulatory circuits that bacteria use to sense and respond to the environment. His other research interests include two-component signaling in E. coli and directed evolution of signaling circuits. Dr. Goulian received his PhD from Harvard University. </div><br />
</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/MillerJordan_Penn-Fellow-e1317190345626.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Dr. Jordan S. Miller</b><br />
<br><br><br />
Dr. Jordan S. Miller is a postdoctoral fellow at the University of Pennsylvania in Dr. Christopher S. Chen's Tissue Microfabrication Laboratory in the Department of Bioengineering. He is currently also a board member of Hive76 and before his time at Penn, he was a developer at RepRap and and an associate at PTV Sciences. Dr. Miller received a PhD from Rice University and earned his undergraduate degree from MIT.<br />
</div></div><br />
<div class="und">Acknowledgements</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/DaphneNg-e1317243791521-224x300.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Daphne Ng</b><br />
<br><br><br />
Daphne Ng is a PhD candidate at the University of Pennsylvania in Dr. Casim A. Sarkar's Molecular Cell Engineering Laboratory in the Department of Bioengineering. She graduated from Cornell University in 2008 with a B.S. in Chemical Engineering. The 2012 Penn iGEM team would like to thank Daphne for her contribution to the cloning and design process, as well as providing expression and cloning vectors.</div><br />
</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/najafshah.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Najaf A. Shah</b><br />
<br><br><br />
Najaf Shah is a PhD candidate at the University of Pennsylvania in Dr. Casim A. Sarkar's Molecular Cell Engineering Laboratory in the Department of Bioengineering. The 2012 Penn iGEM team would like to thank Najaf for providing input on cloning, input on system design, and access to advanced imaging facilities.</div><br />
</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/board-sevile.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Sevile Mannickarottu</b><br />
<br><br><br />
Sevile Mannickarottu is the Director of Bioengineering Instructional Laboratories at the University of Pennsylvania. Before his current post, he was the Philadelphia Operations Manager for Technology Education Awareness and a Project Electrical Engineer at Lutron Electronics. He received an MLA in Religious Studies and a BSc in Electric Engineering from the University of Pennsylvania and is currently pursuing a PhD in Religious Studies. The 2012 Penn iGEM team would like to thank Sevile for providing his facilities and equipment for use during the project.<br />
</div></div><br />
<div class="und">Contributions</div><br />
<br />
<div class="bigbox"><br />
The Penn iGEM team would also like to thank the following individuals for their significant contributions to the team this year:<br />
<ul style="font-size:17px;padding-left:20px; margin:0 0;"><br />
<li><b>Christopher Fang-Yen</b>, PhD, Assistant Professor of Bioengineering at University of Pennsylvania</li><br />
<li><b>Matthew Lazzara</b>, PhD, Assistant Professor of Bioengineering at University of Pennsylvania</li><br />
<li><b>Dan Cohen</b>, PhD, Postdoctoral Fellow at University of Pennsylvania</li><br />
<li><b>Henry Ma</b>, Engineer at University of Pennsylvania Bioengineering Instructional Laboratories</li><br />
<li><b>Karsticum Computing Inc.</b>, Software development company that was instrumental in the construction of VerifiGEM</li></ul><br />
</div><br />
</div><br />
</body><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/TeamTeam:Penn/Team2012-10-27T03:18:21Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<!-----------------------------------------------------------------------------------------------------><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #000000;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.flpic{float:left; margin:0 0 0 0; width:150px;}<br />
.pic1{width:150px;}<br />
.name{ <br />
font-size:20px;<br />
}<br />
.flleft{ float:left; width:400px; margin:0 0 0 50px;}<br />
</style><br />
<br />
<br />
<div class="all"><br />
<br><br />
<div style="text-align:center;font-size:34px;color:white;"><b>Our Team</b></div><br><br />
<br />
<div class="bigbox"><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/7/7a/Penn-iGEM-2012-Team-Photo.jpg" height="500" width="700" /></div><br><br />
The Penn iGEM 2012 Team consists of 4 undergraduates, 3 advisors, and many others who have provided important contributions along the way. Together, we have learned a lot over the last few months. We have taught ourselves different protocols and developed new standards to synchronize our work. Through our participation in iGEM, we have collaborated to learn more about synthetic biology - from the initial days of cloning constructs to the final days of imaging and analysis. Our idea for spatio-temporal control of drug delivery first originated in late May after weeks of reading papers. Slowly we have been able to piece together different components of the system to help the project materialize to the system it is today. We believe our work in optogenetics and drug delivery has a promising future and we are excited to share our results at the World Championships! <br />
</div><br />
<br />
<br />
<br />
<div class="und">Undergraduates</div><br />
<br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="https://static.igem.org/mediawiki/2012/c/c6/Clark_Park_4.jpg" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Ashwin Amurthur</b><br />
<br><br />
<i>"Now we're rolling"</i><br></div><br />
<br><br />
Ashwin is a sophomore at the University of Pennsylvania studying bioengineering and management. He hopes to attend medical school in the future.<br />
<br><br><br />
<b>Natural lab habitat:</b> On the confocal<br><br />
<b>Best kept lab secret:</b> Never ran a PCR <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/9mbudvt">www.tinyurl.com/9mbudvt</a></b> <br />
</div></div><br />
<br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/283077_1448493822523_1538520160_31353808_6931705_n-e1317115984293-221x300.jpg" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Michael Magaraci</b><br />
<br><i>"Sorry guys, I overslept..."</i><br></div><br><br />
Mike is a senior at the University of Pennsylvania studying bioengineering and management. <br />
<br><br><br />
<b>Natural lab habitat:</b> At the PCR machine<br><br />
<b>Best kept lab secret:</b> <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/9rpe7ap">www.tinyurl.com/9rpe7ap</a></b> <br />
</div></div><br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/Untitled-e1317089819600.png" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Peter Qiao</b><br />
<br><i>"There's nothing mini about my miniprep yields"</i><br></div><br><br />
Peter is a junior at the University of Pennsylvania studying bioengineering. He hopes to pursue an M.D/Ph.D in the future.<br />
<br><br><br />
<b>Natural lab habitat:</b> In the hood passaging cells<Br><br />
<b>Best kept lab secret:</b> Triple ligation <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/8e2zj3l">www.tinyurl.com/8e2zj3l</a></b> <br />
<br />
</div></div><br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/P1000543-e1317095308771-221x300.jpg" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Avin Veerakumar</b><br />
<br><i>"Do you want to sleep or do you want to win?"</i><br></div><br><br />
Avin is a senior at the University of Pennsylvania studying Bioengineering and Management.<br />
<br><br><br />
<b>Natural lab habitat:</b> On the confocal<br><br />
<b>Best kept lab secret:</b> Never ran a PCR <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/9ahlsc4">www.tinyurl.com/9ahlsc4</a></b> <br />
</div></div><br />
<br />
<div class="und">Advisors</div><br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/sarkar_web-e1317090202691.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Dr. Casim A. Sarkar</b><br />
<br><br><br />
Dr. Casim A. Sarkar is the Principal Investigator of the Molecular Cell Engineering Laboratory at the University of Pennsylvania Department of Bioengineering. His research interests include molecular cell engineering, protein engineering, ligand/receptor binding and trafficking, cell signaling and decision making, and computational, synthetic, and systems biology. Dr. Sarkar received a PhD in Chemical Engineering with a minor in Computational Biology at MIT and a BS in Chemical Engineering at the University of Texas at Austin. <br />
</div></div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://www.med.upenn.edu/apps/my/images/faculty_pics/goul2044.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Dr. Mark Goulian</b><br />
<br><br><br />
Dr. Mark Goulian is the Edmund J. and Louise W. Kahn Endowed Term Professor of Biology at the University of Pennsylvania. His research is focused on the regulatory circuits that bacteria use to sense and respond to the environment. His other research interests include two-component signaling in E. coli and directed evolution of signaling circuits. Dr. Goulian received his PhD from Harvard University. </div><br />
</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/MillerJordan_Penn-Fellow-e1317190345626.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Dr. Jordan S. Miller</b><br />
<br><br><br />
Dr. Jordan S. Miller is a postdoctoral fellow at the University of Pennsylvania in Dr. Christopher S. Chen's Tissue Microfabrication Laboratory in the Department of Bioengineering. He is currently also a board member of Hive76 and before his time at Penn, he was a developer at RepRap and and an associate at PTV Sciences. Dr. Miller received a PhD from Rice University and earned his undergraduate degree from MIT.<br />
</div></div><br />
<div class="und">Acknowledgements</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/DaphneNg-e1317243791521-224x300.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Daphne Ng</b><br />
<br><br><br />
Daphne Ng is a PhD candidate at the University of Pennsylvania in Dr. Casim A. Sarkar's Molecular Cell Engineering Laboratory in the Department of Bioengineering. She graduated from Cornell University in 2008 with a B.S. in Chemical Engineering. The 2012 Penn iGEM team would like to thank Daphne for her contribution to the cloning and design process, as well as providing expression and cloning vectors.</div><br />
</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/najafshah.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Najaf A. Shah</b><br />
<br><br><br />
Najaf Shah is a PhD candidate at the University of Pennsylvania in Dr. Casim A. Sarkar's Molecular Cell Engineering Laboratory in the Department of Bioengineering. The 2012 Penn iGEM team would like to thank Najaf for providing input on cloning, input on system design, and access to advanced imaging facilities.</div><br />
</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/board-sevile.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Sevile Mannickarottu</b><br />
<br><br><br />
Sevile Mannickarottu is the Director of Bioengineering Instructional Laboratories at the University of Pennsylvania. Before his current post, he was the Philadelphia Operations Manager for Technology Education Awareness and a Project Electrical Engineer at Lutron Electronics. He received an MLA in Religious Studies and a BSc in Electric Engineering from the University of Pennsylvania and is currently pursuing a PhD in Religious Studies. The 2012 Penn iGEM team would like to thank Sevile for providing his facilities and equipment for use during the project.<br />
</div></div><br />
<div class="und">Contributions</div><br />
<br />
<div class="bigbox"><br />
The Penn iGEM team would also like to thank the following individuals for their significant contributions to the team this year:<br />
<ul style="font-size:17px;padding-left:20px; margin:0 0;"><br />
<li><b>Christopher Fang-Yen</b>, PhD, Assistant Professor of Bioengineering at University of Pennsylvania</li><br />
<li><b>Matthew Lazzara</b>, PhD, Assistant Professor of Bioengineering at University of Pennsylvania</li><br />
<li><b>Dan Cohen</b>, PhD, Postdoctoral Fellow at University of Pennsylvania</li><br />
<li><b>Henry Ma</b>, Engineer at University of Pennsylvania Bioengineering Instructional Laboratories</li><br />
<li><b>Karsticum Computing Inc.</b>, Software development company that was instrumental in the construction of VerifiGEM</li></ul><br />
</div><br />
</div><br />
</body><br />
</html></div>Amurthurhttp://2012.igem.org/Team:Penn/TeamTeam:Penn/Team2012-10-27T03:18:05Z<p>Amurthur: </p>
<hr />
<div>{{:Team:Penn/Template/Site2}}<br />
<html><br />
<!-----------------------------------------------------------------------------------------------------><br />
<style type="text/css"><br />
.all{ width:1000px; margin:0 auto;}<br />
.ImageBorder {border: 3px solid #000000;margin: 0 0 20px 0;}<br />
.und{font-size:20px;color:white; text-align:center; margin:20px 0 20px 0;}<br />
.bigbox{width:916px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px;color:black; font-size:14px; text-align:justify;}<br />
.smallbox{ margin:20px auto; width:600px; padding:40px;background:#d7dce1;border:2px solid #708090;border-radius:10px;-moz-border-radius:10px;-webkit-border-radius:10px; overflow:hidden; text-align:justify;}<br />
.flpic{float:left; margin:0 0 0 0; width:150px;}<br />
.pic1{width:150px;}<br />
.name{ <br />
font-size:20px;<br />
}<br />
.flleft{ float:left; width:400px; margin:0 0 0 50px;}<br />
</style><br />
<br />
<br />
<div class="all"><br />
<br><br />
<div style="text-align:center;font-size:34px;color:white;"><b>Our Team</b></div><br><br />
<br />
<div class="bigbox"><br />
<div align="center"><img src="https://static.igem.org/mediawiki/2012/7/7a/Penn-iGEM-2012-Team-Photo.jpg" height="500" width="700" /></div><br><br />
The Penn iGEM 2012 Team consists of 4 undergraduates, 3 advisors, and many others who have provided important contributions along the way. Together, we have learned a lot over the last few months. We have taught ourselves different protocols and developed new standards to synchronize our work. Through our participation in iGEM, we have collaborated to learn more about synthetic biology - from the initial days of cloning constructs to the final days of imaging and analysis. Our idea for spatio-temporal control of drug delivery first originated in late May after weeks of reading papers. Slowly we have been able to piece together different components of the system to help the project materialize to the system it is today. We believe our work in optogenetics and drug delivery has a promising future and we are excited to share our results at the World Championships! <br />
</div><br />
<br />
<br />
<br />
<div class="und">Undergraduates</div><br />
<br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="https://static.igem.org/mediawiki/2012/c/c6/Clark_Park_4.jpg" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Ashwin Amurthur</b><br />
<br><br />
<i>"Now we're rolling"</i><br></div><br />
<br><br />
Ashwin is a sophomore at the University of Pennsylvania studying bioengineering and management.<br />
<br><br><br />
<b>Natural lab habitat:</b> On the confocal<br><br />
<b>Best kept lab secret:</b> Never ran a PCR <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/9mbudvt">www.tinyurl.com/9mbudvt</a></b> <br />
</div></div><br />
<br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/283077_1448493822523_1538520160_31353808_6931705_n-e1317115984293-221x300.jpg" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Michael Magaraci</b><br />
<br><i>"Sorry guys, I overslept..."</i><br></div><br><br />
Mike is a senior at the University of Pennsylvania studying bioengineering and management. <br />
<br><br><br />
<b>Natural lab habitat:</b> At the PCR machine<br><br />
<b>Best kept lab secret:</b> <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/9rpe7ap">www.tinyurl.com/9rpe7ap</a></b> <br />
</div></div><br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/Untitled-e1317089819600.png" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Peter Qiao</b><br />
<br><i>"There's nothing mini about my miniprep yields"</i><br></div><br><br />
Peter is a junior at the University of Pennsylvania studying bioengineering. He hopes to pursue an M.D/Ph.D in the future.<br />
<br><br><br />
<b>Natural lab habitat:</b> In the hood passaging cells<Br><br />
<b>Best kept lab secret:</b> Triple ligation <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/8e2zj3l">www.tinyurl.com/8e2zj3l</a></b> <br />
<br />
</div></div><br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/P1000543-e1317095308771-221x300.jpg" class="pic1" /></div><div class="flleft"><br />
<div style="text-align:center;"><b class="name">Avin Veerakumar</b><br />
<br><i>"Do you want to sleep or do you want to win?"</i><br></div><br><br />
Avin is a senior at the University of Pennsylvania studying Bioengineering and Management.<br />
<br><br><br />
<b>Natural lab habitat:</b> On the confocal<br><br />
<b>Best kept lab secret:</b> Never ran a PCR <br><br />
<b>Favorite Lab Celebration GIF: <a href="http://tinyurl.com/9ahlsc4">www.tinyurl.com/9ahlsc4</a></b> <br />
</div></div><br />
<br />
<div class="und">Advisors</div><br />
<br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/sarkar_web-e1317090202691.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Dr. Casim A. Sarkar</b><br />
<br><br><br />
Dr. Casim A. Sarkar is the Principal Investigator of the Molecular Cell Engineering Laboratory at the University of Pennsylvania Department of Bioengineering. His research interests include molecular cell engineering, protein engineering, ligand/receptor binding and trafficking, cell signaling and decision making, and computational, synthetic, and systems biology. Dr. Sarkar received a PhD in Chemical Engineering with a minor in Computational Biology at MIT and a BS in Chemical Engineering at the University of Texas at Austin. <br />
</div></div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://www.med.upenn.edu/apps/my/images/faculty_pics/goul2044.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Dr. Mark Goulian</b><br />
<br><br><br />
Dr. Mark Goulian is the Edmund J. and Louise W. Kahn Endowed Term Professor of Biology at the University of Pennsylvania. His research is focused on the regulatory circuits that bacteria use to sense and respond to the environment. His other research interests include two-component signaling in E. coli and directed evolution of signaling circuits. Dr. Goulian received his PhD from Harvard University. </div><br />
</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/MillerJordan_Penn-Fellow-e1317190345626.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Dr. Jordan S. Miller</b><br />
<br><br><br />
Dr. Jordan S. Miller is a postdoctoral fellow at the University of Pennsylvania in Dr. Christopher S. Chen's Tissue Microfabrication Laboratory in the Department of Bioengineering. He is currently also a board member of Hive76 and before his time at Penn, he was a developer at RepRap and and an associate at PTV Sciences. Dr. Miller received a PhD from Rice University and earned his undergraduate degree from MIT.<br />
</div></div><br />
<div class="und">Acknowledgements</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/DaphneNg-e1317243791521-224x300.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Daphne Ng</b><br />
<br><br><br />
Daphne Ng is a PhD candidate at the University of Pennsylvania in Dr. Casim A. Sarkar's Molecular Cell Engineering Laboratory in the Department of Bioengineering. She graduated from Cornell University in 2008 with a B.S. in Chemical Engineering. The 2012 Penn iGEM team would like to thank Daphne for her contribution to the cloning and design process, as well as providing expression and cloning vectors.</div><br />
</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/najafshah.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Najaf A. Shah</b><br />
<br><br><br />
Najaf Shah is a PhD candidate at the University of Pennsylvania in Dr. Casim A. Sarkar's Molecular Cell Engineering Laboratory in the Department of Bioengineering. The 2012 Penn iGEM team would like to thank Najaf for providing input on cloning, input on system design, and access to advanced imaging facilities.</div><br />
</div><br />
<div class="smallbox"><div class="flpic"><br />
<img src="http://benjaminshyong.com/igem2011/wp-content/uploads/2011/09/board-sevile.jpg" class="pic1" /></div><div class="flleft"><br />
<b class="name">Sevile Mannickarottu</b><br />
<br><br><br />
Sevile Mannickarottu is the Director of Bioengineering Instructional Laboratories at the University of Pennsylvania. Before his current post, he was the Philadelphia Operations Manager for Technology Education Awareness and a Project Electrical Engineer at Lutron Electronics. He received an MLA in Religious Studies and a BSc in Electric Engineering from the University of Pennsylvania and is currently pursuing a PhD in Religious Studies. The 2012 Penn iGEM team would like to thank Sevile for providing his facilities and equipment for use during the project.<br />
</div></div><br />
<div class="und">Contributions</div><br />
<br />
<div class="bigbox"><br />
The Penn iGEM team would also like to thank the following individuals for their significant contributions to the team this year:<br />
<ul style="font-size:17px;padding-left:20px; margin:0 0;"><br />
<li><b>Christopher Fang-Yen</b>, PhD, Assistant Professor of Bioengineering at University of Pennsylvania</li><br />
<li><b>Matthew Lazzara</b>, PhD, Assistant Professor of Bioengineering at University of Pennsylvania</li><br />
<li><b>Dan Cohen</b>, PhD, Postdoctoral Fellow at University of Pennsylvania</li><br />
<li><b>Henry Ma</b>, Engineer at University of Pennsylvania Bioengineering Instructional Laboratories</li><br />
<li><b>Karsticum Computing Inc.</b>, Software development company that was instrumental in the construction of VerifiGEM</li></ul><br />
</div><br />
</div><br />
</body><br />
</html></div>Amurthur