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Team:Paris Bettencourt/Achievements
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Creation of a new category in the part registry : Semantic containment. The aim of this category is to let people improving each part by adding for instance other amber mutations to existing part to increase the containment. | Creation of a new category in the part registry : Semantic containment. The aim of this category is to let people improving each part by adding for instance other amber mutations to existing part to increase the containment. | ||
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'''Suicide system''' | '''Suicide system''' | ||
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Part K914001 is well characterized and provides immunity to sensitive cells against the Colicin E2 activity protein, but is leaky. Part K914002 is promoterless and allows users to easily plug in the appropriate promoter for their desired purpose. | Part K914001 is well characterized and provides immunity to sensitive cells against the Colicin E2 activity protein, but is leaky. Part K914002 is promoterless and allows users to easily plug in the appropriate promoter for their desired purpose. | ||
* Creation of a new category in the part registry : [http://partsregistry.org/Biosafety XNase]. The aim of this category is to provide users with DNase/RNase parts that can be used for improved kill switches featuring the degradation of genomic material. | * Creation of a new category in the part registry : [http://partsregistry.org/Biosafety XNase]. The aim of this category is to provide users with DNase/RNase parts that can be used for improved kill switches featuring the degradation of genomic material. | ||
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'''MAGE''' | '''MAGE''' | ||
Revision as of 01:30, 27 September 2012
- Main
- Team
- Project
- Achievements
- Human Practice
- Safety
- Notebook
- Attributions
Overview
Delay system
Semantic containment
Restriction enzyme system
MAGE
Suicide system
Encapsulation
Synthetic import domain
Safety Questions
Safety Assessment
Achievements of all the different modules
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Semantic containment Aims Creating a semantic containment system to prevent gene expression in natural organisms Characterize the system Use this system in all genes of the system, the critical genes first (e.g. colicin) System An amber codon (stop codon) embedded in protein genes to prevent their expression and an amber suppressor system in our genetically engineered bacteria Achievements : Construction and characterization of 2 biobricks : K914000 : PLac-supD-T : tRNA amber suppressor K914009 : P1003* Ser133->Amber Codon : kanamycin gene resistance with 1 amber mutation Both part were well characterized and works well. For the second parts, we show that as expected, one mutation is quite leaky, although it works qualitatively, but one mutation is not enough if we want to release such parts in nature. Other reasons emphasize this observation, notably the weakness of being at one mutation to recover the protein functionality. Creation of a new category in the part registry : Semantic containment. The aim of this category is to let people improving each part by adding for instance other amber mutations to existing part to increase the containment. |
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Suicide system Aims : Implement a kill-switch that features population-level suicide and complete genome degradation. System : A synthetic toxin-anti-toxin system based on the wild type Colicin E2 operon. Achievements : We showed that Colicin E2 cells induce cell death in sensitive populations, and that these sensitive populations can be protected by providing them with our engineered immunity protein.
Part K914001 is well characterized and provides immunity to sensitive cells against the Colicin E2 activity protein, but is leaky. Part K914002 is promoterless and allows users to easily plug in the appropriate promoter for their desired purpose.
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MAGE
Aim
Removal of four FseI restriction sites from E. coli MG1655 genome.
Experimental System
Using multiplex automated genome engineering (MAGE) - a technique capable of editing the genome by making small changes in existing genomic sequences.
Achievements
Proof of concept by introducing a stop codon in the middle of the lacZ gene
Synthetic Import Domain Aim
Creation of a novel protein import mechanism in bacteria.
Experimental System
Exploit the natural Colicin import domain fused to any protein at will, dubbed here: "Synthetic Import Domain".
Achievements
- Construction of colicin-like toxin by fusing Colicin E2 based "Synthetic Import Domain" with RNAse domain of colicin D
- Constructon of FseI, I-SceI, LuxR active fragment, LacZ alpha fragment, PyrF and T7 RNA polymerase fused to the two types of "Synthetic Import Domains" from Colicin E2 and Colicin D
- Proof of concept with LacZ alpha fragment fused to "Synthetic Import Domain" from Colicin D
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Human Practice
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