Team:Amsterdam

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<p><b>Introduction</b><br>
 
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Prokaryotes have been selected through evolutionary processes for accurate sensing
 
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and acting upon their living environments. This bacterial versatility can be used by
 
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us, humans, to sense the environments in places we have trouble reaching. Maybe we
 
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would want to measure the conditions (e.g. nutrient availability, toxicity, pathogen
 
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presence, light) somewhere deep under the ground, perhaps we would want to noninvasively
 
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scan for biomarkers in diseased tissue in our bodies. The classical way
 
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to make a bacteria tell us whether a certain event has happened is to link it to the
 
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transcription of fluorescent proteins. This however requires constant monitoring and
 
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maintenance in order to get an idea of the time-variation of the studied system. Could
 
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we make the cell ‘remember’ what it has sensed and when so we can leave it alone for a
 
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while and make it report back to us later?<br><br>
 
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Meet <b>'Me C0l1'</b>, a methylation based celullar log, which uses the naturally occurring phenomenon
 
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of <b>DNA methylation</b> to robustly store signals it has sensed in its environment. The
 
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Amsterdam iGEM 2012 team, consisting of six students, will dedicate the summer to
 
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the realization of this innovative and ambitious plan. This novel storage mechanism,
 
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redesignating an evolutionarly designed tested and proven principle for novel purposes,
 
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could be linked to any of the many biological sensors that are available in the DNA
 
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parts registry. E. memo therefore holds great promise as a detect & store–system for
 
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experimental and industrial purposes.<br><br>
 
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Just storing whether certain signals have been sensed by the cell is only half of the story
 
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however. The proposed memory mechanism would be a form of volatile memory,
 
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of which the traces slowly dissappear as the E. memo-population keeps proliferating.
 
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This is because methylation-patterns are not transferred to the progeny in eukaryotes.
 
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We can use this our advantage. The most exciting part of our project would be to infer
 
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when a signal has been sensed from the percentage of bits that is methylated, which
 
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slowly decreases as the cells keep proliferating. This way, we won’t just store whether
 
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a certain signal has occured; we will also know when it happened.<br><br>
 
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<b>Molecular mechanism</b><br>
 
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In short, we will introduce a site-specific methyltransferase into the iGEM default
 
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chassis organism E. coli, that will only be active/transcribed when the measured signal
 
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is encountered by the logbook-cell. The activated methyltransferase will then move
 
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over to a plasmid region we’ve termed the bit and append a methyl-group to it. By
 
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1
 
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linking the methyltransferase to a Zinc-Finger, its site-specificity is greatly increased,
 
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reducing the amount of undesired background methylation events to a minimum.
 
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Furthermore, by slowing down the cell replication cycle of the cells, we can increase
 
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the span of time we can use to do measurements on.</p>
 
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Revision as of 00:49, 8 August 2012