Difference between revisions of "Team:Fudan-TSI/Notebook"

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                    <p style="width: 100%;text-align: center;font-size: 24px"><span class="white-text">Notebook</span></p>
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                    <ul class="collapsible expandable">
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                        <li>Team: Fudan-TSI</li>
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<li><div class="collapsible-header">Project</div>
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        <li><a href="/Team:Fudan-TSI/Description">Background</a></li>
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                     <h2 style="margin: 0;padding: 10px 0;">Project Summary</h2>
 
                     <h2 style="margin: 0;padding: 10px 0;">Project Summary</h2>
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                     <p style="margin: 0">abstract abstract abstract abstract abstract abstract abstract abstract abstract abstract abstract abstract abstract abstract abstract abstract due sep 5 due sep 5 due sep 5 due sep 5 due sep 5 due sep 5 due sep 5 due sep 5 due sep 5 due sep 5 due sep 5 due sep 5
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                     <p style="margin: 0">Mutation library generation is critical for biological and medical research, but current methods cannot mutate a specific sequence continuously without manual intervention. Here we present a toolbox for <i>in vivo</i> continuous mutation library construction. First, the target DNA is transcribed into RNA. Next, our reverse transcriptase reverts RNA into cDNA, during which the target is randomly mutated by enhanced error-prone reverse transcription. Finally, the mutated version replaces the original sequence through recombination. These steps will be carried out iteratively, generating a random mutation library of the target with high efficiency as mutations accumulate along with bacterial growth. Our toolbox is orthogonal and provides a wide range of applications among various species. R-Evolution could mutate coding sequences and regulatory sequences, which enables the <i>in vivo</i> evolution of individual proteins or multiple targets at a time, promotes high-throughput research, and serves as a foundational advance to synthetic biology.
 
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                         </a><a href="http://www.yfc.cn/en/" target="_blank"><img class="col s3 m6 l3" style="padding: 0.15rem 0.9rem;" alt="Yunfeng Capital" src="https://static.igem.org/mediawiki/2018/e/e2/T--Fudan--yunfengLogo.png">
 
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                             <h3 class="col s12" style="text-align: left; color: rgba(255, 255, 255, 0.8); font-size: 18px">Repeated Evolution in vivo</h3>
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                             <h3 class="col s12" style="text-align: left; color: rgba(255, 255, 255, 0.8); font-size:12px">R-Evolution: an <i>in vivo</i> sequence-specific toolbox for continuous mutagenesis</h3>
 
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                                     <span><a href="/Team:Fudan-TSI/Results">Results</a></span>
 
                                     <span><a href="/Team:Fudan-TSI/Results">Results</a></span>
 
                                     <ul>
 
                                     <ul>

Revision as of 05:05, 15 September 2019

<script src="https://code.jquery.com/jquery-1.11.3.min.js"></script> 2019 Team:Fudan-TSI Notebook

Our notebook

Here we present a main outline of our progress throughout the year.

Our notebook

Here we present a main outline of our progress throughout the year.

Monthly Highlights

April

  • First functional SynNotch constructed
  • Developed a simple testification method of SynNotch—shaking beads linked with antigen
  • Transcription factor and promotor pairs constructed and tested

May

  • SynNotch leakage problem detection
  • Successfully constructed and tested the first mutated version of SynNotch
  • Started constructing composite promoters for logic gates

June

  • SynNotch with various extra- and intra-cellular domain construction
  • SynNotch and composite promoter verification in cell

August

  • Batch construction of plasmids used in “amplifier” and “combiner” layers for all sixteen logic
  • Continual construction and testification of various mutation versions of SynNotch

September

  • Successfully constructed double-stable cell line of two SynNotch receptors
  • Finished constructing plasmids used in logic gates

October

  • Parts construction
  • All sixteen logic testification in cell
project summary

Project Summary

Mutation library generation is critical for biological and medical research, but current methods cannot mutate a specific sequence continuously without manual intervention. Here we present a toolbox for in vivo continuous mutation library construction. First, the target DNA is transcribed into RNA. Next, our reverse transcriptase reverts RNA into cDNA, during which the target is randomly mutated by enhanced error-prone reverse transcription. Finally, the mutated version replaces the original sequence through recombination. These steps will be carried out iteratively, generating a random mutation library of the target with high efficiency as mutations accumulate along with bacterial growth. Our toolbox is orthogonal and provides a wide range of applications among various species. R-Evolution could mutate coding sequences and regulatory sequences, which enables the in vivo evolution of individual proteins or multiple targets at a time, promotes high-throughput research, and serves as a foundational advance to synthetic biology.