Team:Paris Bettencourt/Overview

From 2012.igem.org

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<a class="thumbnail" href="#thumb"><img src="/wiki/images/6/6f/DelaySystem.png" width="60px" border="0" /><span><img src="/wiki/images/b/b9/Delay1PB12.gif" width="500px" /><br /><div id="txtOV">Delay system: This module will permit our bacteria to perform its function during X amount of time  before triggering the degradation and suicide of the bacteria.
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<a class="thumbnail" href="#thumb"><img src="/wiki/images/6/6f/DelaySystem.png" width="60px" border="0" /><span><img src="/wiki/images/b/b9/Delay1PB12.gif" width="500px" /><br /><div id="txtOV">
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Principle: In presence of Arabinose, LacI  is produced repressing the expression of a restriction enzyme. Once Arabinose is not anymore present, the LacI repressor concentration goes down with dilution and degradation leading to the expression of the restriction enzyme.
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1) Physical containment: Our cells will be encapsulated with Arabinose and LB in order to avoid release of DNA/cells and to permit growth
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2)Delay system: In presence of Arabinose, LacI  is produced repressing the expression of a restriction enzyme. Once Arabinose is not anymore present, the LacI repressor concentration goes down with dilution and degradation leading to the expression of the restriction enzyme.
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<a class="thumbnail" href="#thumb"><img src="/wiki/images/5/5c/RestrictionSystem.png" width="60px" border="0" /><span><img src="/wiki/images/9/97/Restriction2PB12.gif" width="500px" /><br /><div id="txtOV">Restriction Enzyme system system.</div></span></a>
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<a class="thumbnail" href="#thumb"><img src="/wiki/images/5/5c/RestrictionSystem.png" width="60px" border="0" /><span><img src="/wiki/images/9/97/Restriction2PB12.gif" width="500px" /><br /><div id="txtOV">
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1)Physical containment: Our cells will be encapsulated with Arabinose and LB in order to avoid release of DNA/cells and to permit growth
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2)Delay system: In presence of Arabinose, LacI  is produced repressing the expression of a restriction enzyme. Once Arabinose is not anymore present, the LacI repressor concentration goes down with dilution and degradation leading to the expression of the restriction enzyme.
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3)Restriction Enzyme system: The restriction enzyme destroy a plasmid carrying an anti-toxin.</div></span></a>
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<a class="thumbnail" href="#thumb"><img src="/wiki/images/7/78/ToxinAntitoxin.png" width="60px" border="0" /><span><img src="/wiki/images/8/8f/Toxin3aPB12.gif" width="500px"/><br /><div id="txtOV">The suicide system is a variation of a toxin-anti-toxin system, and consists of a lethal DNA cleaving protein which is bound with high affinity by the anti-toxin immunity protein. The anti-toxin plasmid carrying the immunity protein, when combined with the restriction enzyme system, is degraded on a tunable delay. Once the anti-toxin plasmid degrades, the toxin kills the cell, its neighbors, and eliminates extracellular DNA via its DNase activity.</div></span></a>
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<a class="thumbnail" href="#thumb"><img src="/wiki/images/7/78/ToxinAntitoxin.png" width="60px" border="0" /><span><img src="/wiki/images/8/8f/Toxin3aPB12.gif" width="500px"/><br /><div id="txtOV"> 1)Physical containment: Our cells will be encapsulated with Arabinose and LB in order to avoid release of DNA/cells and to permit growth
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2)Delay system: In presence of Arabinose, LacI  is produced repressing the expression of a restriction enzyme. Once Arabinose is not anymore present, the LacI repressor concentration goes down with dilution and degradation leading to the expression of the restriction enzyme.
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3)Restriction Enzyme system: The restriction enzyme destroy a plasmid carrying an anti-toxin cassette
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4)Suicide system: Once the anti-toxin is below a given threshold the toxin is not anymore titrated , the toxin kills the cell, its neighbors, and eliminates extracellular DNA via its DNase activity.</div></span></a>
<a class="thumbnail" href="#thumb"><img src="https://static.igem.org/mediawiki/2012/1/1c/SemanticContainment.png" width="60px" border="0" /><span><img src="/wiki/images/8/8f/Toxin3aPB12.gif" width="500px" /><br /><div id="txtOV">Semantic Containment.</div></span></a>
<a class="thumbnail" href="#thumb"><img src="https://static.igem.org/mediawiki/2012/1/1c/SemanticContainment.png" width="60px" border="0" /><span><img src="/wiki/images/8/8f/Toxin3aPB12.gif" width="500px" /><br /><div id="txtOV">Semantic Containment.</div></span></a>
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<a class="thumbnail" href="#thumb"><img src="https://static.igem.org/mediawiki/2012/1/1c/SemanticContainment.png" width="60px" border="0" /><span><img src="/wiki/images/8/8f/Toxin3aPB12.gif" width="500px" /><br /><div id="txtOV">Toxin/anti-Toxin system.</div></span></a>
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<a class="thumbnail" href="#thumb"><img src="https://static.igem.org/mediawiki/2012/1/1c/SemanticContainment.png" width="60px" border="0" /><span><img src="/wiki/images/8/8f/Toxin3aPB12.gif" width="500px" /><br /><div id="txtOV">1) Physical containment: Our cells will be encapsulated with Arabinose and LB in order to avoid release of DNA/cells and to permit growth </div></span></a>
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Revision as of 20:27, 26 September 2012


iGEM Paris Bettencourt 2012


1) Physical containment: Our cells will be encapsulated with Arabinose and LB in order to avoid release of DNA/cells and to permit growth 2)Delay system: In presence of Arabinose, LacI is produced repressing the expression of a restriction enzyme. Once Arabinose is not anymore present, the LacI repressor concentration goes down with dilution and degradation leading to the expression of the restriction enzyme.

1)Physical containment: Our cells will be encapsulated with Arabinose and LB in order to avoid release of DNA/cells and to permit growth 2)Delay system: In presence of Arabinose, LacI is produced repressing the expression of a restriction enzyme. Once Arabinose is not anymore present, the LacI repressor concentration goes down with dilution and degradation leading to the expression of the restriction enzyme. 3)Restriction Enzyme system: The restriction enzyme destroy a plasmid carrying an anti-toxin.

1)Physical containment: Our cells will be encapsulated with Arabinose and LB in order to avoid release of DNA/cells and to permit growth 2)Delay system: In presence of Arabinose, LacI is produced repressing the expression of a restriction enzyme. Once Arabinose is not anymore present, the LacI repressor concentration goes down with dilution and degradation leading to the expression of the restriction enzyme. 3)Restriction Enzyme system: The restriction enzyme destroy a plasmid carrying an anti-toxin cassette 4)Suicide system: Once the anti-toxin is below a given threshold the toxin is not anymore titrated , the toxin kills the cell, its neighbors, and eliminates extracellular DNA via its DNase activity.

Semantic Containment.

1) Physical containment: Our cells will be encapsulated with Arabinose and LB in order to avoid release of DNA/cells and to permit growth
















































Project Overview

Contents

Objectives

Our project aims to:

  • Raise the issue of biosafety, and advocate the discerning use of biosafety circuits in future iGEM projects as a requirement
  • Evaluate the risk of HGT in different SynBio applications
  • Develop a new, improved containment system to expand the range of environments where GEOs can be used safely.

To do so, we:

  • Engaged the general public and scientific community through debate
  • Raised the question about how we can regulate this practices
  • Compiled a parts page of safety circuits in the registry
  • Relied on three levels of containment :
    1. Physical containment with alginate capsules
    2. Semantic containment using an amber suppressor system
    3. An improved killswitch featuring delayed population-level suicide through complete genome degradation.

We strived to make our system as robust against mutations as possible.

General recommandation for a good killswitch device

  • bla
  • blo
  • blu

Step 1

Step 2

Step 2.

Step 3

Step 3.












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