Team:Cambridge/Quiescence

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=Quiescence=
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{{:Team:Cambridge/Templates/headerbar|colour=#fb5c2b|title=Shelved project: Quiescence}}
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This Project seeks to design a device that can be used to switch cells into a non-growing, but metabolically active state. A system like this would be highly desirable, as it could be used to increase the efficiency of expression of plasmid genes. It could also help to reduce the frequency of undesirable mutations and provide a supplementary bio-safety mechanism.
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This Project seeks to design a device that can be used to switch cells into a non-growing, but metabolically active state. A system like this would be highly desirable, as it could be used to increase the efficiency of expression of plasmid genes. Quiescence would be a useful safeguard against loss of plasmids, which can cause the culture to be overgrown by plasmid-free bacteria. It could also help to reduce the frequency of undesirable mutations and provide a supplementary bio-safety mechanism, if escaping bacteria were unable to replicate.
=='''R'''egulator of '''c'''ell '''d'''ivision RNA==
=='''R'''egulator of '''c'''ell '''d'''ivision RNA==
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A new approach to making a quiescence switch would be to control the functionality of the folded RNA instead of its transcription, which proved to be problematic. Ideally the bacteria would grow in the lab or a vat in the presence of a ligand but would stop growing without dying, as soon as the ligand is removed. Thus if the engineered bacteria escaped into the wild they would stop growing very quickly as the ligand is diluted.  
A new approach to making a quiescence switch would be to control the functionality of the folded RNA instead of its transcription, which proved to be problematic. Ideally the bacteria would grow in the lab or a vat in the presence of a ligand but would stop growing without dying, as soon as the ligand is removed. Thus if the engineered bacteria escaped into the wild they would stop growing very quickly as the ligand is diluted.  
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We hope to achieve this by rationally designing a gene for an RNA that folds into a non-functional shape. The structure will include a hammerhead ribozyme domain (see [[http://sage.ucsc.edu/scottlab/reprints/2006_Scott_Cell_pdf/2006_Scott_Cell.pdf Martick and Scott 2006]]) that will hopefully cleave itself in vivo to release the functional Rcd RNA. In order to control this cleavage, the final construct would include an aptamer, which is an RNA structure able to bind a specific compound. This would create an allosteric ribozyme.
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We hope to achieve this by rationally designing a gene for an RNA that folds into a non-functional shape. The structure will include a hammerhead ribozyme domain (see [http://sage.ucsc.edu/scottlab/reprints/2006_Scott_Cell_pdf/2006_Scott_Cell.pdf Martick and Scott 2006]) that will hopefully cleave itself in vivo to release the functional Rcd RNA. In order to control this cleavage, the final construct would include an aptamer, which is an RNA structure able to bind a specific compound. This would create an allosteric ribozyme.
The project would involve synthesising different constructs (which are all mercifully short) and testing their functionality and sensitivity. The result would be a fairly short, but quite versatile BioBrick.
The project would involve synthesising different constructs (which are all mercifully short) and testing their functionality and sensitivity. The result would be a fairly short, but quite versatile BioBrick.
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==Intellectual Property==
==Intellectual Property==
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Using Rcd in the h-ns mutant to induce quiescence in E.coli is the Intellectual Property of Cambridge Microbial technologies. Dr Summers has been very supportive and we are going to continue the project. However, the legal issues have not been completely resolved yet. We still need to talk to Robert Hulme, the co-holder of the patent.  
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Using Rcd in the h-ns mutant to induce quiescence in E.coli is the Intellectual Property of Cambridge Microbial technologies. Dr Summers and Dr Hulme have been very supportive.
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==Further notes==
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Rough notes are available [https://2010.igem.org/Team:Cambridge/Quiescence_Notes here].
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Latest revision as of 23:55, 16 August 2010