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Team:Paris Bettencourt/Delay
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As an alternative trigger to the «collective suicide» presented in the overview of the project, we tried to build a system in which the toxin would be cloned without the Immunity protein, in order to avoid any possible resistance to the toxin due to a non-degraded plasmid. Such a design comes with several difficulties : | As an alternative trigger to the «collective suicide» presented in the overview of the project, we tried to build a system in which the toxin would be cloned without the Immunity protein, in order to avoid any possible resistance to the toxin due to a non-degraded plasmid. Such a design comes with several difficulties : | ||
| - | + | *we need a '''very strong repression''' of the expression of the toxin. A leaky expression would harm the cells, and one of the risks is that mutants which do not produce the toxin arise and become selected. | |
| - | + | *we need to be able to have a '''control''' on the delay, in order for the cells to trigger the safety mechanism only once they have performed their function. | |
| - | + | *we need to be sure that the safety system will be triggered at one point. '''We prioritize the robustness of the delay over its timespan'''. | |
===Design=== | ===Design=== | ||
| + | |||
| + | In order to achieve our goal, we started form the construct of Yokobayashi ''et al.'' [ref] described below. | ||
==Experiments and results== | ==Experiments and results== | ||
Revision as of 10:03, 24 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 AssessmentContents |
Overview
This part of the design will trigger the DNA degradation mechanism. To do so, we will use two different approaches. The first one, that we called "simple delay system", will be a way to trigger a restriction enzyme, which will cut the plasmid carrying the antitoxin, and thus let the cell sensitive to the toxin produced on the chromosome. In the "sRNA delay system, we use a different strategy. Here, the toxin would be cloned without the Immunity protein, in order to avoid any possible resistance to the toxin due to a non-degraded plasmid. To ensure that the toxin will not be produced while the cells are performing their function, we will use a stationary phase promoter and sRNA repression induced by arabinose.
Simple delay system
Objectives
Design
sRNA delay system
Objectives
As an alternative trigger to the «collective suicide» presented in the overview of the project, we tried to build a system in which the toxin would be cloned without the Immunity protein, in order to avoid any possible resistance to the toxin due to a non-degraded plasmid. Such a design comes with several difficulties :
- we need a very strong repression of the expression of the toxin. A leaky expression would harm the cells, and one of the risks is that mutants which do not produce the toxin arise and become selected.
- we need to be able to have a control on the delay, in order for the cells to trigger the safety mechanism only once they have performed their function.
- we need to be sure that the safety system will be triggered at one point. We prioritize the robustness of the delay over its timespan.
Design
In order to achieve our goal, we started form the construct of Yokobayashi et al. [ref] described below.
Experiments and results
Characterisation of X
Experimental setup
Describe the experiment
Results
Present your results
Testing of the system
Experimental setup
Describe the experiment
Results
Present your results
