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 provide an easy-to-use BLACK BOX, 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! :)
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The BREAKTHROUGH we made: an easy-to-use BLACK BOX, a versatile platform for in vivo optimization of metabolic pathways
