Team:SJTU-BioX-Shanghai

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Revision as of 22:12, 27 September 2013

Abstract

How to regulate an entire metabolic pathway in vivo, delicately, accurately and conveniently, simultaneously controlling the expression of several genes? And how to optimize metabolic fluxes so as to maximize desired products? For decades, these questions remain haunting to “bioengineers”(especially synthetic biologists) who would like to have certain metabolites produced in live cells. The difficulty is again raised up when target genes endogenously reside on the genome, or have been implemented into the genome.

So this year, our team, SJTU-BioX-Shanghai, is dedicated to solve the problem. By integrating CRISPRi, the newly developed expression interference tool, with three light sensors (namely red light sensor, green light sensor and blue light sensor), we expect to provide a versatile platform on which researchers are able to quantitatively adjust the expression of any three enzymes in whatever pathway – to change the target, simply change the small guide RNA (sgRNA)!

In addition, with the luminous device and accompanying software, one could simply enter metabolomics data (catalytic rates of related enzymes) into our User Interface. The software will automatically conduct flux balance analysis (FBA), giving out suggestions for optimal expression value of different enzymes. Then, the expression value would be converted into intensity of lights that are finally to be exerted on cell cultures. It is -- just convenient! :)

Luminous System provide us a fascinating tool to adjust expression on genomic level.

We can optimize a metabolic system by typing different numbers on computer!

Achievements

The breakthrough we made: Redefinition of scaffold in Synthetic Biology by recruiting E.coli’s inner membrane as a natural two-dimensional scaffold.

The system we built: 6 membrane proteins orderly organized on the inner membrane of E.coli, the efficiency of which has been proved by fluorescence complementation assay and biosynthesis experiment.

The device we created – Membrane Accelerator: A universal tool that serves to accelerate biochemical reactions in E.coli; Rate of fatty acids synthesis was increased by 24 fold compared to wild-type E.coli and 9 fold compared to that with overexpressed cytoplasmic enzymes.

device we created – Membrane Rudder: A universal tool used to dynamically and artificially control biochemical reactions in E.coli; the direction of Violacein and Deoxyviolacein synthetic pathway was successfully switched.

New direction we proposed: The application of scaffold system in accelerating biodegradation pathway using our Membrane Accelerator.

Parts we submitted: 42 well-characterized parts that could either be used directly or serve as a universal tool readily for potential scientific or engineering use.

A club we established – BioCraft: The headquarter of our human practice programs, having come a long way in propagandizing Synthetic Biology and iGEM. Warmly-received activities have been held in and outside the campus. Several celebrities in different fields have shown support for us, laying a cornerstone for our future development.

Sponsors


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