Team:Aberdeen Scotland

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<h1>Project Abstract</h1>
<h1>Project Abstract</h1>
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Our team aimed to engineer a genetic toggle switch that allows yeast to express either green or cyan fluorescent protein (GFP & CFP) mutually exclusively. This is regulated at the translational level which provides a more efficient and quicker response than previous switches regulated at the transcriptional level, bypassing mRNA synthesis and export. This is a novel mechanism previously untried. Our biology team successfully tested the RNA-binding protein/fluorescent protein expression in yeast, at the single cell and population levels; provided the modelling team parameters obtained from experimentation and research; and sequenced BioBricks fused to GFP and CFP in yeast expression cassettes. Using this data our modelling team successfully created a deterministic and a stochastic model of how we expected the toggle switch to behave and found that the maximum success rate for our original set of parameters was 0.96%, but through theoretical modifications an optimal parameter set predicted a maximum success rate of 51.27%. Unfortunately time constraints prevented implementation of these adjustments to the system, a procedure we would recommend for further research.
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Our team engineered a novel genetic toggle switch regulated at the translational level that allowed yeast to express either green or cyan fluorescent protein mutually exclusively. We successfully constructed a fusion of mRNA binding protein and fluorescent protein in yeast characterized using microscopy, a fluorometer and FACS analysis of single cells or whole populations. These results along with parameters from literature were used to predict via deterministic and stochastic models that the probability of successful bistability for our switch was 0.96%, but this could be improved theoretically to a maximum of 51.27% by limiting the range of variation of the most sensitive parameters. The models also implied that co-operativity of binding of the mRNA binding protein to its mRNA stem loop was theoretically essential for generating switch-like behaviour. These results suggest that a genetic toggle switch regulated at the translational level is a viable and novel engineering concept applicable to medicinal, environmental and technological problems.
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Revision as of 23:06, 18 September 2010

University of Aberdeen - ayeSwitch - iGEM 2010

Project Abstract

Our team engineered a novel genetic toggle switch regulated at the translational level that allowed yeast to express either green or cyan fluorescent protein mutually exclusively. We successfully constructed a fusion of mRNA binding protein and fluorescent protein in yeast characterized using microscopy, a fluorometer and FACS analysis of single cells or whole populations. These results along with parameters from literature were used to predict via deterministic and stochastic models that the probability of successful bistability for our switch was 0.96%, but this could be improved theoretically to a maximum of 51.27% by limiting the range of variation of the most sensitive parameters. The models also implied that co-operativity of binding of the mRNA binding protein to its mRNA stem loop was theoretically essential for generating switch-like behaviour. These results suggest that a genetic toggle switch regulated at the translational level is a viable and novel engineering concept applicable to medicinal, environmental and technological problems.


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