Team:Peking/Project/Overview

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  <h3 id="title1">Achievements</h3>
  <h3 id="title1">Achievements</h3>
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This summer, we have<br />
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Here is a list of our achievements from this summer.<br /><BR>
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1. rationally constructed a hypersensitive sensor of luminance – what we call the <i>Luminesensor</i>. <br />
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1. We helped four other iGEM teams by sharing DNA materials, characterizing their parts and modeling!  Click <a href="/Team:Peking/HumanPractice/Outreach/Collaboration"><i> Here</i></a><br /><br />
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2. Primarily, the sensor was designed by fusing a blue-light-sensing protein domain from <i>Neurospora</i> with a DNA binding domain LexA from <i>E. coli</i>. Following this, protein structure inspection and kinetic simulations were conducted to rationally perform optimization. Amazingly, the <i>Luminesensor</i> proved to be as sensitive to sense natural light and even bioluminescence. With this sensor, spatiotemporal control of cellular behavior, such as phototaxis, high-resolution 2-D and 3-D bio-printing using dim light, and even utilizing the luminescence of an iPad were shown to be quite possible. What’s more, we successfully implemented cell-cell communication using light for the very first time in synthetic biology, which will prove to be of great importance for biotechnological use.  
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2. We outlined in detail a new approach of human pratice called "Sowing Tomorrow's Synthetic Biologists"! Click <a href="/Team:Peking/HumanPractice/Sowing"><i> Here</i></a><br /><br />
 +
3. We presented all fresh iGEMers with a collection and praise of historic iGEM projects to share and learn from each other!Click <a href="/Team:Peking/HumanPractice/Review"><i> Here</i></a><br /><br />
 +
4. We submitted 11 high quality and well-characterized standard biobricks! Click <a href="/Team:Peking/DataPage"><i> Here</i></a><br /><br />
 +
5. We improved the ease-of-use of "Lux Brick" that primarily constructed by Cambridge iGEM 2010 and carefully characterized its dynamics of functionality!  Click <a href="/wiki/index.php?title=Team:Peking/Project/Communication/Results#title2"><i> Here</i></a> or <a href="http://partsregistry.org/Part:BBa_K819008">MainPage</a><br /><br />
 +
6. We successfully implemented spatiotemporal control of cellular behavior, such as high-resolution 2-D and 3-D bio-printing using dim light, and even utilizing the luminescence of an iPad!  Click <a href="/Team:Peking/Project/3D/2D"><i> Here</i></a><br /><br />
 +
7. We successfully implemented cell-cell communication using bioluminescence for the very first time in synthetic biology!  Click <a href="/Team:Peking/Project/Communication/Results"><i> Here</i></a><br /><br />
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We deserve a <b>Gold Medal Prize</b>.<br />
  </p>
  </p>
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  <h3 id="title2">1. Luminesensor <br /> -- An Ultrasensitive Transcription Factor</h3>
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  <h3 id="title2">1. The <i>Luminesensor</i> <br> -- An Ultrasensitive Light Sensor</h3>
  <p>
  <p>
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Based on the statistical data, Peking iGEM has concluded that an ultrasensitive transcription factor may circumvent the serious issues of current optogenetics methods, e.g. cytotoxicy, narrow dynamic range, and dependency on laser and exogenous chromophore.
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Based on comprehensive literature reviews, our team has concluded that an ultra-sensitive light-sensing light-off system which constructed by 3 our ECUST members from Yang lab, here we called the <i>Luminesensor</i> may circumvent serious issues of current optogenetic methods, e.g. cytotoxicity, narrow dynamic range, and dependency on laser and exogenous chromophores.  
   <br /> <br />
   <br /> <br />
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Inspired by the basic design principle of past optogenetics methods, a light sensitive domain was fused with a functional domain. To meet the basic criteria of not needing exogenous chromophores, we chose the smallest LOV (light, oxygen, or voltage) protein—Vivid (VVD) from Neurospora as the light sensitive domain. Based on the dimerization mechanism of VVD, the DNA binding domain, LexA from <i>E. coli</i>, was fused to the N-terminal of VVD in order to couple the light signals with the repression of gene expression (Figure 1). The target gene expression is repressed by the <i>Luminesensor</i> when illuminated by blue light (440~480nm).
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</p>
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<p>
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Amazingly, the <i>Luminesensor</i> proved to be sensitive to natural light and even bioluminescence. Moreover, the <i>Luminesensor</i> is able to respond to light signals with broad illuminance scales (Figure 1).
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   <img src="/wiki/images/4/4f/Peking2012_LuminesensorMechanism.jpg" alt="" style="width:500px;" />
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   <img src="/wiki/images/d/d0/Peking2012_IlluminanceScales.png" alt="" style="width:600px;" />
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Figure 1. The basic design of Luminesensor.
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Figure 1. The broad illuminance scales to which <i>Luminesensor</i> could respond to.
   </p>
   </p>
   </div>
   </div>
  </div>
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</div>
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<h3 id="title3">2. Synthetic Biology in 2D and 3D <br /> -- A Practical Application</h3>
  <p>
  <p>
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Based on this fundamental design, protein structure inspection and kinetic simulations were conducted to rationally perform optimization. (If you would like to know the details of the <i>Luminesensor</i> design, please see the <a href="/Team:Peking/Project/Luminesensor/Design">Design page</a>.)
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Few optogenetics methods have been applied to practical applications until now. To fully demonstrate spatiotemporally precise manipulation of cellular behavior using our <i>Luminesensor</i>, our team has deeply explored the applications  in 2D and 3D bio-printing.
-
  <br /> <br />
+
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Due to the consideration of minimal influence on endogenous components, a special mutant of LexA was used to ensure the orthogonality with the endogenous LexA protein. The orthogonal <i>Luminesensor</i> proved to work very well in the common <i>E. coli</i> strain without LexA deletion (data in <a href="/Team:Peking/Project/Luminesensor/Characterization">Characterization</a>).
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  <br /> <br />
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Consistent with the results of kinetic simulation (details in <a href="/Team:Peking/Modeling/Luminesensor">Modeling</a>), mutagenesis based on previous kinetic data of VVD mutation largely improved the property of the <i>Luminesensor</i>, now with an amazing dynamic range reaching 300(data in <a href="/Team:Peking/Project/Luminesensor/Characterization">Characterization</a>).
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   <br /> <br />
   <br /> <br />
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The high sensitivity of the VVD protein is preserved when coupled with the LexA DNA binding domain. Amazingly, the <i>Luminesensor</i> proves to be sensitive to natural light and even bioluminescence (data in <a href="/Team:Peking/Project/Luminesensor/Characterization">Characterization</a>). Such high sensitivity allows the <i>Luminesensor</i> to respond to light signals with broad illuminance scales (Figure 2).
+
High resolution images could be easily obtained by 2D bioprinting on a bacterial lawn bearing the <i>Luminesensor</i> (Figure 2). (more on <a href="/Team:Peking/Project/3D/2D">2D Printing</a>).
  </p>
  </p>
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   <img src="/wiki/images/d/d0/Peking2012_IlluminanceScales.png" alt="" style="width:600px;" />
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   <img src="/wiki/images/c/ca/Peking2012_PrintParagraph.jpg" alt="" style="width:500px;" />
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Figure 2. The broad illuminance scales that the <i>Luminesensor</i> could respond to.
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Figure 2. High resolution of 2D bioprinting.
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   </div>
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  <p>
  <p>
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Unsatisfied yet with the properties of the <i>Luminesensor</i>, we also wants to red-shift the absorption spectrum of VVD through molecular docking (see <a href="/Team:Peking/Modeling/Channel">Modeling</a>) and mutation of VVD (see <a href="/Team:Peking/Modeling/Luminesensor">Extension</a>), which would largely extend the future application of the <i>Luminesensor</i>, e.g. a multicolor <i>Luminesensor</i> control system.
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What's more, sharp images could be printed using an Apple iPad as light source (see <a href="/Team:Peking/Project/3D/2D">iPrinting</a>), which indicates that our <i>Luminesensor</i> could serve as the interface between biological systems and electrical devices.
 +
  <br /> <br />
 +
A very promising application of the <i>Luminesensor</i> is 3D printing, using light signals of spatially high resolution. Three dimensional images could be elegantly printed into media mixed with bacteria expressing the <i>Luminesensor</i> (see <a href="/Team:Peking/Project/3D/3D">3D Printing</a>). A new design strategy of 3D printer using holographic technology instead of layer-by-layer printing, was also proposed. With such a power tool, 3D printing regarding biological material has endless potentials for medical use (crazy ideas in <a href="/Team:Peking/Project/3D/Future">Future</a>).
  </p>
  </p>
</div>
</div>
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  <h3 id="title3">2. Syn Bio in 2D and 3D <br /> -- An Industrial Application <br /> of Luminesensor</h3>
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  <h3 id="title4">3. Cell-Cell Light Communication <br /> -- A Novel Strategy of Signaling</h3>
  <p>
  <p>
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Few optogenetics methods have been applied to industrial use until now. To fully demonstrate the precise spatiotemporal manipulation of light and the bright future of optogenetics, Peking iGEM has deep explored the application of the <i>Luminesensor</i> in 2D and 3D bioprinting methods.
+
Cell-cell communication based on quorum sensing systems, e.g. AHL, has been largely utilized by synthetic biologists to construct gene circuits with complex functions, e.g. pattern formation and edge detection. However, the diffusion and saturation of quorum sensing chemicals limit the delivery of communication signals, which in turn requires high spatiotemporal resolution and long range interaction.
   <br /> <br />
   <br /> <br />
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High resolution image could be obtained by 2D bioprinting on the plate mixed with <i>Luminesensor</i> (Figure 3), using easily constructed devices (more on <a href="/Team:Peking/Project/3D/2D">2D Printing</a>).
+
The ultrasensitivity of the <i>Luminesensor</i> encouraged our team to explore the possibility of cell-cell light communication (Figure 3). By employing bacteria expressing the lux operon from <i>V. fisheri</i> as "light senders" and those bearing the <i>Luminesensor</i> as "light receiver", we successfully demonstrated, for the first time ever, that light communication between cells could be achieved beyond direct physical contact (see <a href="/Team:Peking/Project/Communication/Results">video</a> in Results). Through measuring the dynamics of light communication, it was proved that bioluminescence was sufficient to trigger the response of the <i>Luminesensor</i> (data in <a href="/Team:Peking/Project/Communication/Results">Results</a>).
  </p>
  </p>
  <div class="floatC">
  <div class="floatC">
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   <img src="/wiki/images/c/ca/Peking2012_PrintParagraph.jpg" alt="" style="width:500px;" />
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   <img src="/wiki/images/9/9c/Peking2012_overview_achieve_communication.png" alt="" style="width:500px;" />
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   <p class="description">
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Figure 3. High resolution 2D bioprinting.
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Figure 3. The concept of cell-cell communication through light.
   </p>
   </p>
   </div>
   </div>
  </div>
  </div>
  <p>
  <p>
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Even more amazingly, sharp images could be printed using the Retina Display of an Apple iPad (see <a href="/Team:Peking/Project/3D/2D">iPrinting</a>), which serves as the interface between biological systems with electrical devices.
+
Our Light-Off system could be upgraded into Light-On system through simple design (details in <a href="/Team:Peking/Project/Communication/Design#title3">Design</a>), which enables us to achieve more flexible cell-cell light communication (creative ideas in <a href="/Team:Peking/Project/Communication/Future">Future</a>).  
-
  <br /> <br />
+
-
A very promising area of the industrial applications of the <i>Luminesensor</i> is 3D printing, due to the high spatiotemporal specificity of light signals. 3D images could be elegantly printed into the plate mixed with the <i>Luminesensor</i> (see <a href="/Team:Peking/Project/3D/3D">3D Printing</a>). A new design idea of 3D printer, using holographic technology instead of layer-by-layer printing. This could raise a revolution in the 3D printing world. With such a power tool, 3D printing with biological material has endless potentials in the medical field (crazy ideas on <a href="/Team:Peking/Project/3D/Future">Future</a>).
+
  </p>
  </p>
</div>
</div>
<div class="PKU_context floatR">
<div class="PKU_context floatR">
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  <h3 id="title4">3. Cell-Cell Light Communication <br /> -- A New Generation of Optogenetics</h3>
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  <h3 id="title5">4. Phototaxis <br /> --Controlling Cell Motion by Light</h3>
  <p>
  <p>
-
Cell-cell communication based on quorum sensing systems, e.g. AHL, has been largely utilized by synthetic biologists to construct gene circuits with complex functions, e.g. pattern formation and edge detection. However, the diffusion and saturation of quorum sensing chemicals limit the delivery of communication signals, which in turn requires high spatiotemporal resolution and long range interaction.
+
What makes <i>Luminesensor</i> outstanding is not only its ultra-sensitivity, but also the particularly high dynamic range (data on <a href="/Team:Peking/Project/Luminesensor/Characterization#title3">Characterization</a>). Such switch-like property makes <i>Luminesensor</i> a very reliable and beneficial module in synthetic biology. Similar to the light-controlled animal behavior in neuroscience, by coupling <i>Luminesensor</i> with endogenous systems, it is possible to integrate the light signal with cell motion.  
-
  <br /> <br />
+
-
The ultrasensitivity of the <i>Luminesensor</i> encouraged our team to explore the possibility of cell-cell light communication (Figure 4). By employing the lux operon from V. fisheri as the light sender and the <i>Luminesensor</i> as the light receiver, we successfully demonstrated, for the first time ever, that light communication between cells could be achieved beyond direct physical contact (see <a href="/Team:Peking/Project/Communication/Results">video</a> in Results). Through measuring the difference of the reporter gene expression, it was proved that the light of the lux operon was sufficient to trigger the response of the <i>Luminesensor</i> (data in <a href="/Team:Peking/Project/Communication/Results">Results</a>).
+
-
  <br /> <br />
+
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A complete light communication system with positive and negative control was constructed by modifying the Light-Off system to Light-On system (details in <a href="/Team:Peking/Project/Communication/Design">Design</a>), which enables us to achieve more complex functions based on cell-cell light communication (creative ideas in <a href="/Team:Peking/Project/Communication/Future">Future</a>).  
+
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  <img src="/wiki/images/c/c1/Peking2012_Home_Phototaxis.jpg" alt="" style="width:500px;" />
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<h3 id="title5">4. Phototaxis <br /> -- Cell Motion Controlled by Light</h3>
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  <div>
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  <p class="description">
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Figure 4. The concept of phototaxis behavior.
 +
  </p>
 +
  </div>
 +
</div>
  <p>
  <p>
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What makes the <i>Luminesensor</i> outstanding is not only its ultra-sensitivity, but also the particularly high dynamic range (data on <a href="/Team:Peking/Project/Luminesensor/Characterization">Characterization</a>). Such switch-like property makes the <i>Luminesensor</i> a very reliable and beneficial module in synthetic biology. Similar to the light-controlled animal behavior in neuroscience, by coupling the <i>Luminesensor</i> with endogenous systems, it is possible to integrate the light signal with the native response to control biological behavior on a higher level, e.g. cell motion.
+
"Phototactic" bacteria can be built by reprogramming the chemotaxis system in <i>E. coli</i> through light with the <i>Luminesensor</i> (Figure 4). By controlling the expression of the CheZ protein with light, the tumbling frequency is coupled to intensity of light signals (details in <a href="/Team:Peking/Project/Phototaxis/Design">Design</a>). On the border of the light and dark fields, motility ability of the cells in a single colony on the two sides is sufficient to result in an uneven colony (see <a href="/Team:Peking/Project/Phototaxis/Demonstration">Demonstration</a>). Light-controlled cellular motion indeed has very promising environmental and medical applications for the future, e.g. the delivery of drugs to target concerns.
-
  <br /> <br />
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The 2012 Peking iGEM has successfully built "Phototatic" bacteria by programming the chemotaxis system in <i>E. coli</i> through light with the <i>Luminesensor</i> (Figure 5). By controlling the expression level of the cheZ protein with light, the tumbling frequency is coupled to the intensity of light signals (details in <a href="/Team:Peking/Project/Phototaxis/Design">Design</a>). On the border of the light and dark fields on the plate, the motility difference of the cells in a single colony on the two sides is sufficient to form an uneven colony (see <a href="/Team:Peking/Project/Phototaxis/Demonstration">Demonstration</a>). The light-controlled cell motion that we have achieved has a very promising environmental and medical application for the future, e.g. the delivery of drugs to target concerns.
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  </p>
  </p>
</div>
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</html>{{Template:Peking2012_Color_Epilogue}}
</html>{{Template:Peking2012_Color_Epilogue}}

Latest revision as of 07:48, 16 October 2012

Achievements

Here is a list of our achievements from this summer.

1. We helped four other iGEM teams by sharing DNA materials, characterizing their parts and modeling! Click Here

2. We outlined in detail a new approach of human pratice called "Sowing Tomorrow's Synthetic Biologists"! Click Here

3. We presented all fresh iGEMers with a collection and praise of historic iGEM projects to share and learn from each other!Click Here

4. We submitted 11 high quality and well-characterized standard biobricks! Click Here

5. We improved the ease-of-use of "Lux Brick" that primarily constructed by Cambridge iGEM 2010 and carefully characterized its dynamics of functionality! Click Here or MainPage

6. We successfully implemented spatiotemporal control of cellular behavior, such as high-resolution 2-D and 3-D bio-printing using dim light, and even utilizing the luminescence of an iPad! Click Here

7. We successfully implemented cell-cell communication using bioluminescence for the very first time in synthetic biology! Click Here

We deserve a Gold Medal Prize.

1. The Luminesensor
-- An Ultrasensitive Light Sensor

Based on comprehensive literature reviews, our team has concluded that an ultra-sensitive light-sensing light-off system which constructed by 3 our ECUST members from Yang lab, here we called the Luminesensor may circumvent serious issues of current optogenetic methods, e.g. cytotoxicity, narrow dynamic range, and dependency on laser and exogenous chromophores.

Amazingly, the Luminesensor proved to be sensitive to natural light and even bioluminescence. Moreover, the Luminesensor is able to respond to light signals with broad illuminance scales (Figure 1).

Figure 1. The broad illuminance scales to which Luminesensor could respond to.

2. Synthetic Biology in 2D and 3D
-- A Practical Application

Few optogenetics methods have been applied to practical applications until now. To fully demonstrate spatiotemporally precise manipulation of cellular behavior using our Luminesensor, our team has deeply explored the applications in 2D and 3D bio-printing.

High resolution images could be easily obtained by 2D bioprinting on a bacterial lawn bearing the Luminesensor (Figure 2). (more on 2D Printing).

Figure 2. High resolution of 2D bioprinting.

What's more, sharp images could be printed using an Apple iPad as light source (see iPrinting), which indicates that our Luminesensor could serve as the interface between biological systems and electrical devices.

A very promising application of the Luminesensor is 3D printing, using light signals of spatially high resolution. Three dimensional images could be elegantly printed into media mixed with bacteria expressing the Luminesensor (see 3D Printing). A new design strategy of 3D printer using holographic technology instead of layer-by-layer printing, was also proposed. With such a power tool, 3D printing regarding biological material has endless potentials for medical use (crazy ideas in Future).

3. Cell-Cell Light Communication
-- A Novel Strategy of Signaling

Cell-cell communication based on quorum sensing systems, e.g. AHL, has been largely utilized by synthetic biologists to construct gene circuits with complex functions, e.g. pattern formation and edge detection. However, the diffusion and saturation of quorum sensing chemicals limit the delivery of communication signals, which in turn requires high spatiotemporal resolution and long range interaction.

The ultrasensitivity of the Luminesensor encouraged our team to explore the possibility of cell-cell light communication (Figure 3). By employing bacteria expressing the lux operon from V. fisheri as "light senders" and those bearing the Luminesensor as "light receiver", we successfully demonstrated, for the first time ever, that light communication between cells could be achieved beyond direct physical contact (see video in Results). Through measuring the dynamics of light communication, it was proved that bioluminescence was sufficient to trigger the response of the Luminesensor (data in Results).

Figure 3. The concept of cell-cell communication through light.

Our Light-Off system could be upgraded into Light-On system through simple design (details in Design), which enables us to achieve more flexible cell-cell light communication (creative ideas in Future).

4. Phototaxis
--Controlling Cell Motion by Light

What makes Luminesensor outstanding is not only its ultra-sensitivity, but also the particularly high dynamic range (data on Characterization). Such switch-like property makes Luminesensor a very reliable and beneficial module in synthetic biology. Similar to the light-controlled animal behavior in neuroscience, by coupling Luminesensor with endogenous systems, it is possible to integrate the light signal with cell motion.

Figure 4. The concept of phototaxis behavior.

"Phototactic" bacteria can be built by reprogramming the chemotaxis system in E. coli through light with the Luminesensor (Figure 4). By controlling the expression of the CheZ protein with light, the tumbling frequency is coupled to intensity of light signals (details in Design). On the border of the light and dark fields, motility ability of the cells in a single colony on the two sides is sufficient to result in an uneven colony (see Demonstration). Light-controlled cellular motion indeed has very promising environmental and medical applications for the future, e.g. the delivery of drugs to target concerns.

  • Totop Totop