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| − | <a href="https://2016.igem.org/"><img src="https://static.igem.org/mediawiki/2014/0/09/Aalto_Helsinki_Logov_iGEM.png" class="img-responsive igem-logo"></a>
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| − | <h1>Clamp Down on Crosstalk<br>
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| − | <small>Alverno_CA</small></h1>
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| − | <center>Reducing Noise in Multi-Gene Synthetic Biology Circuits</center>
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| − | <a href="#Abstract">
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| − | <a href="http://i-see-faces.deviantart.com/">Photo © Tanja Maria</a>
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| − | <div class="link" id="Abstract"></div>
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| − | <h2>Clamping Down on Crosstalk:<br>
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| − | <small>How can we keep genes from interfering with each other in synthetic DNA circuits?</small></h2>
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| − | <center><h3>About this Project</h3></center>
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| − | Building complex biological systems with many genes requires isolating genes. Active genes can cause nearby DNA to become supercoiled, <br>leading to unpredictable behavior of synthetic biology systems. We will test if DNA clamps (made from DNA-binding proteins) placed between <br>genes can stop this interference. If this project succeeds, it will allow bioengineers to build more predictable genetic circuits.</center>
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| − | <a href="https://youtu.be/sFfD-AivQw4"><img src="https://static.igem.org/mediawiki/2016/5/55/Videothumb.png" class="img-responsive smallerimg"></img></a>
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| − | <center>Watch our introduction video here.</center>
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| − | <img src="https://static.igem.org/mediawiki/2014/7/7e/Aalto_Helsinki_Logot_Oma.png" class="img-responsive center-block omalogot">
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| − | <center><h3>What is the context of this research?</h3></center>
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| − | Programming cells often requires building "circuits" of several genes<br> together on the same piece of DNA. Bioengineers have observed that <br>when two genes are placed next to each other, they often unexpectedly interfere with each other's<br> expression in an unexpectedly orientation-dependent manner.
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| − | Nobody knows with certainty what causes this genetic crosstalk, but one promising<br> theory involves DNA supercoiling. The transcription of DNA into RNA, the transcription process introduces supercoils, similar to kinks in a tightly-wound phone <br> cord. Supercoils directly affect the expression of genes, turning them on or off depending on the direction of the supercoil.</center>
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| − | <center><h3>What is the significance of this project?</h3></center>
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| − | If successful, a DNA-binding, gene- isolating clamp could be used in any multi-gene circuit assembly, making multi-gene assemblies more predictable and their assembly much more efficient.<br>
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| − | This is particularly relevant when engineering metabolic pathways to produce chemicals<br> like methanol, insulin, or antibiotics, where circuits of many genes are routinely constructed. The physical layout of these circuits can unpredictably affect production of<br> the desired output by several orders of magnitude, so large engineered metabolic pathways must typically be hand-tuned or have many configurations screened for activity. By<br> making gene expression more predictable, our results could greatly improve the predictability (and, therefore, designability)<br> of large gene circuits for metabolic engineering.</center>
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| − | <center><h3>What are the goals of the project?</h3></center>
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| − | We will first build several plasmids (circular pieces of DNA) that <br>demonstrate cross-talk between genes. These plasmids <br>will consist of genes for two different fluorescent proteins (green fluorescent protein and red fluorescent protein) next to each other, in different orientations. <br>We expect to see differences in the relative expression of the two genes depending on how they are arranged and oriented, and we will quantify this effect.<br>
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| − | Next, we will try several strategies for removing these differences, including adding extra base pairs of spacing between the two genes and adding DNA "clamps" <br> made from DNA-binding repressor proteins between the two genes. We will again quantify the effects of cross-talk between genes, <br> which will hopefully be ameliorated by our additions.</center>
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