Team:Paris Bettencourt/Project/Trojan Horse

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    SABOTAGE
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<div id="grouptitle">Trojan Horse Project</div>
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    <h2>Overview </h2>
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<h2> Introduction </h2>
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<p>Antibiotic resistance is wide spreads in nature and is a major problem especially in the attempt to cure diseases such as tuberculosis.
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Tuberculosis is an infectious disease that typically attacks lungs and which is caused by a mycobacterium called ''Mycobacterium Tuberculosis''. One third of the world's population is thought to have been infected by the mycobacterium. Moreover, in the past years, several strains of the bacterium carrying antibiotics resistance and therefor complicated to treat have appeared.<br>
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<h2> Objectives = make an antibiotic resistant population sensitive to antibiotics </h2>
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<h1>Design a silencing device</h1>
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<h1>Design a genetic element that spread in a population</h1>
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<h2>Design</h2>
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<h1>A silencing system : sRNA </h1>
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<h1>A vector : Phage  </h1>
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<h1>A non counter selected system Phagemid system</h1>
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<h1>Sequential method to kill</h1>
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<h2>Results</h2>
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<h1>Characterization of the phagemid system</h1>
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<i>Experimental set up </i>
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<i>Results</i>
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The treatment against tuberculosis usually takes a lot of time, around six months, and reacquire a lots of drugs partially due to the fact that they are a lot of strains resistant to various antibiotics. It's a huge health problem notably in developing countries. Our project aims to facilitate the treatment of the disease by making it easier to kill the antibiotics resistant strains of the mycobacterium.<br>
 
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Our idea is to embed a synthetic sRNA on a selfish DNA element, for example a phage, that spreads in a bacterial population and to use this construct to select against unwanted genes, in this case antibiotic resistance.<br>
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The synthetic sRNA is a small RNA (24 bp) designed to bind to the first 24 base pairs of the mRNA of an antibiotic gene. We will target chloramphenicol resistance gene and / resistance gene for our proof of concept, using strain MG1655 of E.coli. Once the sRNA has bound, the ribosomes are unable to bind to the RNA which is then not translated into a protein. The cell is not producing the antibiotic resistance protein anymore and is easily killable using antibiotics.<br>
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We also want to work on a construct in which we can repress a LacZ gene to be able to quantify the effectiveness of our synthetic sRNA construct.<br>
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<h1>Sequential killing</h1>
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<i>Experimental set up </i>
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We aim to test several constructs, integrating the resistance or/and the synthetic sRNA construct in plasmid or/and in the chromosome of the bacterium as wild bacteria can have resistance both on chromosome and plasmid.<br>
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<i>Results </i>
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<img src="https://static.igem.org/mediawiki/2013/d/d7/PB_TH_Killingatdifferentconcentration%24.png" width="350px" height="200px"/>
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Starring: Helena, Clovis, Vincent, Aude, Anne</p>
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<h1>Generalization: results for Kan</h1>
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<h1>Origin of resistance </h1>
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<h2>Conclusion and perspectives</h2>
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<h1>Several sRNA on the same plasmid : </h1>
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<h1>Building a library of sRNA : </h1>
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<h1>Possible Applications : </h1>
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Latest revision as of 15:24, 3 October 2013

<body>
SABOTAGE

Overview

Objectives = make an antibiotic resistant population sensitive to antibiotics

Design a silencing device

Design a genetic element that spread in a population

Design

A silencing system : sRNA

A vector : Phage

A non counter selected system Phagemid system



Sequential method to kill

Results

Characterization of the phagemid system

Experimental set up Results


Sequential killing

Experimental set up Results

Generalization: results for Kan

Origin of resistance

Conclusion and perspectives

Several sRNA on the same plasmid :

Building a library of sRNA :

Possible Applications :

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
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