Difference between revisions of "Team:IIT Delhi/standardpage introduction"

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<h1 style="font-size:20px;"><b>Blueprint of our project</b></h1></br>
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If people want to drive a system to work for them, even if the system is a simple one, simply keep pushing the system toward the goal may not be the best shoot. For example, if a professor is too pushing, his students may, on the contrary, unable to perform their best; if a farmer adding too much fertilizer, the land may be damaged in the long run, etc.</br></br>
 
  
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Biological systems are extremely complex, and the components in the system are intensely interconnected. So in order to exploit the maximum potentiality of a biological system, we'll have to keep the protein or metabolic product production in a desired range. Not too high, as it may hurt the cell or inhibit its growth; either not too low, as it will be economically inefficient. </br></br>
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So how can we reach a desired range of expression? We need to properly combine the transcription and translation initiation elements, just as an recent published Nature article suggested[1]. But that paper just used the throughly studied E.coli expression elements in E.coli. What if we are doing engineering in a non-model organism that we just have data about a handful of expression elements, can we create the elements we need? </br></br>
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Our project proposed a way to employ a limited set of promoters to reach any desired expression level, or even switch between several expression level. </br></br>
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        <a href="index.html">Home</a> <span class="divider">/</span>
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        Team
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<h2>Current Team</h2>
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    <h2>Meet the <strong>Team</strong></h2>
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<center><em>Fig.1 Tandem promoter</em></br></center>
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First, we combined the known promoter into tandem promoter system. We've done experiments and modeling to show how can we use a 0.1 promoter and a 0.3 promoter to reach expression level from 0.1 to the maximum. Please check experiment here and modeling here. </br></br>
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<center><em>Fig.2 Cas9 regulated multistage promoter</em></br></center>
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<center><em>Fig 3. An analog to slide rheostat</em></br></center>
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Then, we made the tandem promoter a “slide rheostat” by using d/aCas9 to regulate it. This enable the tandem promoter to switch between several designable expression level, and become a multistage promoter. This is different from the normal regulated promoter that usually has only two stage: on and off (Fig.4). To see our experiment about this multistage promoter, please <a href="https://2013.igem.org/Team:WHU-China/modules#tandem_promoter">click here</a>. It's also important to ensure the orthogonality of this multistage promoter. So the off-target tendency of Cas9 is modeled and analyzed by combining the data of six paper about Cas9 off-target. For the modeling result, please <a href="https://2013.igem.org/Team:WHU-China/modelingintro">click here</a>. </br></br>
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<center><em>Fig 4. Bi-stage promoter and multistage promoter</em></br></center>
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<h1><b>Reference</b></h1>
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<em>[1]Mutalik, Vivek K., et al. "Precise and reliable gene expression via standard transcription and translation initiation elements." Nature methods 10.4 (2013): 354-360.</em>
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Latest revision as of 18:34, 11 September 2015

Current Team

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