UPSC SeminarPaul Hudson
Systems Biology, KTH, Stockholm, Sweden
Title: Synthetic and systems biology of cyanobacteria empowered with the CRISPRi tool
Host: Åsa Strand
More information about Paul Hudson's research: https://www.kth.se/profile/huds
Abstract for the talk:
Our group is using the inducible gene-repression tool CRISPRi in Synechocystis for multiple projects in metabolic engineering and systems biology. We have exploited the multiplex capability (6 sgRNAs) of CRISPRi to rapidly screen the effect of eliminating competing pathways on fatty alcohol production. In this case, we observed regulatory “compensation” when repressing genes around the acyl-ACP node. An inducible CRISPRi allows controlled repression of essential genes, and we have used repression of citrate synthase to arrest growth and direct flux to lactate, where specific productivities are increased several fold. I will discuss how sgRNA libraries, combined with droplet-based cell sorting for lactate secretion, can be used to find better strategies for arresting growth while keeping CO2 fixation high. Finally, I will give an example of how we use CRISPRi repression of enzymes to force accumulation of a metabolite, and perform subsequent transcriptomics to uncover metabolite-responsive gene regulation.