Team:SJTU-BioX-Shanghai

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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! :)

The Luminous System provides a fascinating platform for adjusting expression of several genes by shedding lights of different color with different intensity on the cell culture.

Light sensors transform optical signals into biochemical changes and can then realise biological response interference.

We use CRISPRi system as a direct gene regulator.Under the control of light sensors, we expect to get specific output parameters of gene expression levels.

Achievements

The breakthrough we made: The quantitative regulation of gene expression in the level of genome.

The system we built: Assure the effectiveness of the novel system CRISPRi , integration between CRISPRi system and several sensors.

The device we created : An electronic device constructed to measure the parameters of LED.

Helping other teams: Sending parts to Zhejiang University and the Chinese University of Hong Kong, helping improving the modeling of the projects of NTU-Taida and NTU Taiwan in the 2013 conference, help students from Shandong University to construct their own team.

The experiment we did: Having a great summer filled with experiments, and planning to have fun at the Asian Jamboree!.

Parts we submitted: We make components of popular CRISPRi into parts and improve several parts by inserting them into constitutive expression operon.

Human practise: Involvement in an official academic elite cultivation plan supported by national education ministry. Cooperation with corporation to try applying our project into industry. A survey associated with subject cross and a detailed relevant analysis of this survey.

Help Correct a Mistake in a Published Paper: We help an author realize there is a unit mistake in the published paper on JMB when we design our experiment measurement.