"
Team:Paris Bettencourt/Results
From 2013.igem.org
(Difference between revisions)
| Line 63: | Line 63: | ||
<h2>Aims</h2> | <h2>Aims</h2> | ||
<p>To perform an drug screen targeted at the sirA gene from mycobacteria</p> | <p>To perform an drug screen targeted at the sirA gene from mycobacteria</p> | ||
| + | </div> | ||
| + | <div style="clear: both;"></div> | ||
| + | </div> | ||
| + | <h2>Infiltrate</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="" target="_blank">BBa_K1137008 (TDMH)</a></li> | ||
| + | </ul> | ||
| + | </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> | ||
<div style="clear: both;"></div> | <div style="clear: both;"></div> | ||
Revision as of 23:45, 4 October 2013
Detect
Background
CRISPR/Cas systems generate site-specific double strand breaks and have recently be 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.
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.
Centre for Research and Interdisciplinarity (CRI)
Faculty of Medicine Cochin Port-Royal, South wing, 2nd floor
Paris Descartes University
24, rue du Faubourg Saint Jacques
75014 Paris, France
+33 1 44 41 25 22/25
team2013@igem-paris.org
+33 1 44 41 25 22/25
team2013@igem-paris.org 
