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Team:NRP-UEA-Norwich/Project
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Revision as of 15:19, 20 September 2012 by RussellGritton (Talk | contribs)
Please choose the relevant link to view an overview of each project!
Nitric Oxide Sensing & The Hybrid Promoters | The Comparator Circuit | Theoretical Projects
Sensory BioBrick systems have been a large constituent of previous iGEM projects in which teams have combined impressive amounts of logic with limitless creativity in order to produce synthetically engineered organisms with the ability to detect the presence of specific substrates; this was achieved by combining various promoters and reporters to produce novel gene systems of great breadth and depth.
We too have taken a sensory approach to our project and have produced systems involved in the sensation of nitric oxide (NO). Originally we set out to develop a bacterial and mammalian hybrid NO-sensing promoter (which we have achieved); we then looked into ways of quantifying the levels of highly reactive and difficult to measure NO within a system, leading to us producing a novel gene regulation system known as the comparator circuit. Throughout the project we went on to look at theoretical alternate approaches to the gene systems we have produced.
Overall in our project we have produced 6 sensory BioBricks, 2 BioBricks involved in gene regulation, and have further characterised 4 more BioBricks. All 8 of our BioBricks have been submitted to the registry and the 6 sensory BioBricks have been characterised.
Our own hybrid promoter hopes to add to the systems already in the registry by creating a hybrid promoter that combines the bacterial promoter PyeaR and the mammalian CArG element , both of which respond to exogenous nitrogenous species. Combining the two would allow a more modular NO sensor that can be used in mammalian and bacterial cells interchangeably.
. Flexible and has two orientations
. Six biobricks out of it
. Characterised with fluorescent proteins
. Data from flow cytometry, fluorometers, FACS
. Transfected into mammalian cells for further testing
The lack of specificity of the bacterial promoter, pYEAR, used in the hybrid promoter was a pitfall that was always a concern. From this potential problem spawned a potential solution; the Comparator Circuit. This pair of BioBricks are designed to specifically bind to each other while ligated to promoters of overlapping specificity to result in an integrating of the conflicting outputs of the two opposing gene systems.
In essence, what we have created is a pair of antagonistic BioBricks that turned the pair of mRNAs in which they reside into translational repressor molecules when both are transcribed in tandum within a specific chassis of interest.
