Team:Paris Bettencourt/YonatanTest
From 2013.igem.org
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+ | .results { | ||
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+ | } | ||
+ | .biocriks { | ||
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<img src="https://static.igem.org/mediawiki/2013/d/d9/PB_achievementsbanenr.png" width="1100"/> | <img src="https://static.igem.org/mediawiki/2013/d/d9/PB_achievementsbanenr.png" width="1100"/> | ||
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<div id="page"> | <div id="page"> | ||
- | <h2><a href="https://2013.igem.org/Team:Paris_Bettencourt/Project/Detect">Detect</a></h2> | + | <h2><a href="https://2013.igem.org/Team:Paris_Bettencourt/Project/Detect">Detect</a></h2> |
- | <div class="overbox | + | <div class="overbox"> |
- | + | <div class="bkgr"> | |
- | + | <h2>Background</h2> | |
- | + | <p>CRISPR/Cas systems generate site-specific double strand breaks and have recently been used for genome editing. </p> | |
+ | </div> | ||
+ | <div class="results"> | ||
+ | <h2>Results</h2> | ||
+ | <ul> | ||
+ | <li>Successfully cloned gRNA anti-KAN, crRNA anti-KAN, tracrRNA-Cas9 and pRecA-LacZ into Biobrick backbones and therefore generated four new BioBricks. </li> | ||
+ | <li>Testing the new assembly standard for our cloning.</li> | ||
+ | </ul><h2><a href="https://2013.igem.org/Team:Paris_Bettencourt/Assembly_Standard">Assembly Standard</h2> | ||
+ | <p>We offer a new assembly standard. It enables keeping the BioBrick standard while providing the needed tools to perform assembly of several parts in one step. BBG is a fusion of the BioBrick standard cloning and Gibson isothermal assembly.</p> | ||
+ | </div> | ||
+ | <div class="biocriks"> | ||
+ | <h2>BioBricks</h2> | ||
+ | <ol> | ||
+ | <li><a href="http://parts.igem.org/Part:BBa_K1137012" target="_blank">BBa_K1137012 (gRNA anti-KAN)</a></li> | ||
+ | <li><a href="http://parts.igem.org/Part:BBa_K1137013" target="_blank">BBa_K1137013 (crRNA anti-KAN)</a></li> | ||
+ | <li><a href="http://parts.igem.org/Part:BBa_K1137014" target="_blank">BBa_K1137014 (tracrRNA-Cas9)</a></li> | ||
+ | <li><a href="http://parts.igem.org/Part:BBa_K1137015" target="_blank">BBa_K1137015 (pRecA-LacZ)</a></li> | ||
+ | </ol> | ||
+ | </div> | ||
+ | <div style="clear: both;"></div> | ||
+ | <div class="aims"> | ||
+ | <h2>Aims</h2> | ||
+ | <p>Building a genotype sensor based on CRISPR/Cas that reports existance of an antibiotic resistance gene.</p> | ||
+ | </div> | ||
+ | <div style="clear: both;"></div> | ||
</div> | </div> | ||
- | + | <h2><a href="https://2013.igem.org/Team:Paris_Bettencourt/Project/Target">Target</a></h2> | |
- | <h2>Results</h2> | + | <div class="overbox"> |
- | + | <div class="bkgr"> | |
- | + | <h2>Background</h2> | |
- | + | <p>SirA is an essential gene in latent tuberculosis infections</p> | |
- | + | </div> | |
- | + | <div class="results"> | |
+ | <h2>Results</h2> | ||
+ | <ul> | ||
+ | <li>Produced an E. coli strain which relies upon mycobacterial sirA, fprA and fdxA genes to survive in M9 minimal media</li> | ||
+ | <li>Demonstrated that E. coli can survive with mycobacterial sulfite reduction pathway with Flux Balance Analysis</li> | ||
+ | <li>Located drug target sites on sirA as well as identified high structural similarity between cysI and sirA through structural anaylsis</li> | ||
+ | </ul> | ||
+ | <p></p> | ||
+ | </div> | ||
+ | <div class="biocriks"> | ||
+ | <h2>BioBricks</h2> | ||
+ | <ol> | ||
+ | <li><a href="http://parts.igem.org/Part:BBa_K1137000">BBa_K1137000 (SirA)</a></li> | ||
+ | <li><a href="http://parts.igem.org/Part:BBa_K1137001">BBa_K1137001 (FprA)</a></li> | ||
+ | <li><a href="http://parts.igem.org/Part:BBa_K1137002">BBa_K1137002 (FdxA)</a></li> | ||
+ | </ol> | ||
+ | </div> | ||
+ | <div style="clear: both;"></div> | ||
+ | <div class="aims"> | ||
+ | <h2>Aims</h2> | ||
+ | <p>To perform an drug screen targeted at the sirA gene from mycobacteria</p> | ||
+ | </div> | ||
+ | <div style="clear: both;"></div> | ||
</div> | </div> | ||
- | + | ||
- | <div style="clear: both;"></div> | + | <h2><a href="https://2013.igem.org/Team:Paris_Bettencourt/Project/Infiltrate">Infiltrate</a></h2> |
- | + | <div class="overbox"> | |
- | + | <div class="bkgr"> | |
- | + | <h2>Background</h2> | |
+ | <p>Latent tuberculosis persists inside macrophages of the lungs, where it is partially protected from both the host immune system and conventional antibiotics.</p> | ||
+ | </div> | ||
+ | <div class="results"> | ||
+ | <h2>Results</h2> | ||
+ | <ul> | ||
+ | <li>We expressed the enzyme Trehalose Dimycolate Hydrolase (TDMH) in <i>E.coli</i> and showed that it is highly toxic to mycobacteria in culture.</li> | ||
+ | <li>We expressed the lysteriolyin O (LLO) gene in <i>E. coli</i> and showed that it is capable of entering the macrophage cytosol.</li> | ||
+ | <li>We co-infected macrophages with both mycobacteria and our engineered <i>E. coli</i> to characterize the resulting phagocytosis and killing.</li> | ||
+ | </ul> | ||
+ | <p></p> | ||
+ | </div> | ||
+ | <div class="biocriks"> | ||
+ | <h2>BioBricks</h2> | ||
+ | <ol> | ||
+ | <li><a href="http://parts.igem.org/Part:BBa_K1137008" target="_blank">BBa_K1137008 (TDMH)</a></li> | ||
+ | </ol> | ||
+ | </div> | ||
+ | <div style="clear: both;"></div> | ||
+ | <div class="aims"> | ||
+ | <h2>Aim</h2> | ||
+ | <p>To create an <i>E. coli</i> strain capable of entering the macrophage cytosol and delivering a lytic enzyme to kill mycobacteria.</p> | ||
+ | </div> | ||
+ | <div style="clear: both;"></div> | ||
</div> | </div> | ||
- | + | <h2><a href="https://2013.igem.org/Team:Paris_Bettencourt/Project/Sabotage">Sabotage</a></h2> | |
- | + | <div class="overbox"> | |
- | <h2> | + | <div class="bkgr"> |
- | <ol> | + | <h2>Background</h2> |
- | + | <p>One of the main concern about tuberculosis today is the emergence of antibiotic resistant strain</p> | |
- | + | </div> | |
- | + | <div class="results"> | |
- | + | <h2>Results</h2> | |
- | </ | + | <ul> |
+ | <li>Construction and characterization of phagemids coding for small RNA targeting antibiotic resistance proteins</li> | ||
+ | <li>successful conversion of antibiotic resistant population of E. coli to a sensitive state</li> | ||
+ | </ul> | ||
+ | <p></p> | ||
+ | </div> | ||
+ | <div class="biocriks"> | ||
+ | <h2>BioBricks</h2> | ||
+ | <ol> | ||
+ | <li><a href="http://parts.igem.org/Part:BBa_K1137009" target="_blank">BBa_K1137009 (sRNA anti Kan) Characterized </a></li> | ||
+ | <li><a href="http://parts.igem.org/Part:BBa_K1137010" target="_blank">BBa_K1137010 (sRNA anti Cm) Characterized</a></li> | ||
+ | <li><a href="http://parts.igem.org/Part:BBa_K1137011" target="_blank">BBa_K1137011 (sRNA anti Lac) </a></li> | ||
+ | </ol> | ||
+ | </div> | ||
+ | <div style="clear: both;"></div> | ||
+ | <div class="aims"> | ||
+ | <h2>Aims</h2> | ||
+ | <p>Our objective is to make an antibiotic-resistant bacterial population sensitive again to those same antibiotics.</p> | ||
+ | </div> | ||
+ | <div style="clear: both;"></div> | ||
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</html> | </html> | ||
<!-- ########## Don't edit below ########## --> | <!-- ########## Don't edit below ########## --> | ||
{{:Team:Paris_Bettencourt/footer}} | {{:Team:Paris_Bettencourt/footer}} |
Revision as of 08:30, 25 October 2013
![](https://static.igem.org/mediawiki/2013/d/d9/PB_achievementsbanenr.png)
Detect
Background
CRISPR/Cas systems generate site-specific double strand breaks and have recently been used for genome editing.
Results
- Successfully cloned gRNA anti-KAN, crRNA anti-KAN, tracrRNA-Cas9 and pRecA-LacZ into Biobrick backbones and therefore generated four new BioBricks.
- Testing the new assembly standard for our cloning.
Assembly Standard
We offer a new assembly standard. It enables keeping the BioBrick standard while providing the needed tools to perform assembly of several parts in one step. BBG is a fusion of the BioBrick standard cloning and Gibson isothermal assembly.
BioBricks
Aims
Building a genotype sensor based on CRISPR/Cas that reports existance of an antibiotic resistance gene.
Target
Background
SirA is an essential gene in latent tuberculosis infections
Results
- Produced an E. coli strain which relies upon mycobacterial sirA, fprA and fdxA genes to survive in M9 minimal media
- Demonstrated that E. coli can survive with mycobacterial sulfite reduction pathway with Flux Balance Analysis
- Located drug target sites on sirA as well as identified high structural similarity between cysI and sirA through structural anaylsis
Aims
To perform an drug screen targeted at the sirA gene from mycobacteria
Infiltrate
Background
Latent tuberculosis persists inside macrophages of the lungs, where it is partially protected from both the host immune system and conventional antibiotics.
Results
- We expressed the enzyme Trehalose Dimycolate Hydrolase (TDMH) in E.coli and showed that it is highly toxic to mycobacteria in culture.
- We expressed the lysteriolyin O (LLO) gene in E. coli and showed that it is capable of entering the macrophage cytosol.
- We co-infected macrophages with both mycobacteria and our engineered E. coli to characterize the resulting phagocytosis and killing.
BioBricks
Aim
To create an E. coli strain capable of entering the macrophage cytosol and delivering a lytic enzyme to kill mycobacteria.
Sabotage
Background
One of the main concern about tuberculosis today is the emergence of antibiotic resistant strain
Results
- Construction and characterization of phagemids coding for small RNA targeting antibiotic resistance proteins
- successful conversion of antibiotic resistant population of E. coli to a sensitive state
BioBricks
Aims
Our objective is to make an antibiotic-resistant bacterial population sensitive again to those same antibiotics.