Team:Paris Bettencourt/Overview

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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 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

Story telling: An example of applications for our project

Imagine a farmer that would like to know how much fertilizer is in his field. We provide him cells with nitrate biosensor (AgrEcoli) encapsulated with arabinose. He spray them in his field wait for 12h and check if bids are glowing. Once the result collected, we don't want that any synthetic organism or DNA be released and potentially transfer to a soil organism. That's why once arabinose is completely degraded, the delay system will trigger the degradation of any synthetic DNA followed by the death of our organism.

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.

Step 1

Step 2

Step 2.

Step 3

Step 3.