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Professor Yu-Chen Hu
Chairman, Department of Chemical Engineering, National Tsing Hua University, Taiwan
Title of the talk: CRISPR-Cas9 for the Engineering of Microorganisms and Production of Chemicals
Cyanobacteria hold promise as a cell factory for producing biofuels and bio-derived chemicals, but genome engineering of cyanobacteria such as Synechococcus elongatus PCC 7942 poses challenges because of their oligoploidy nature and long-term instability of the introduced gene. CRISPR-Cas9 is a newly developed RNA-guided genome editing system, yet its application for cyanobacteria engineering has yet to be reported. Here we demonstrated that CRISPR-Cas9 system can effectively trigger programmable double strand break (DSB) at the chromosome of PCC 7942 and provoke cell death. With the co-transformation of template plasmid harboring the gene cassette and flanking homology arms, CRISPR-Cas9-mediated DSB enabled precise gene integration, ameliorated the homologous recombination efficiency and allowed the use of lower amount of template DNA and shorter homology arms. The CRISPR-Cas9-induced cell death imposed selective pressure and enhanced the chance of concomitant integration of gene cassettes into all chromosomes of PCC 7942, hence accelerating the process of obtaining homogeneous and stable recombinant strains. We further explored the feasibility of engineering cyanobacteria by CRISPR-Cas9-assisted simultaneous glgc knock-out and gltA/ppc knock-in, which improved the succinate titer to 435.0±35.0 μmg/L, an ≈ 11-fold increase when compared with that of the wild-type cells. These data altogether justify the use of CRISPR-Cas9 for genome engineering and manipulation of metabolic pathways in cyanobacteria.
Dr. Yu-Chen Hu received his BS degree in Chemical Engineering from National Taiwan University (1992) and earned his PhD degree in Chemical Engineering from University of Maryland (USA) in 1999. He worked as a post-doc at the National Institutes of Health from 1999 to 2000 and returned to the Department of Chemical Engineering, National Tsing Hua University, Taiwan, in 2000. Dr. Hu’s main research interests include vaccine development, gene therapy, tissue engineering, cancer therapy and synthetic biology. Dr. Hu has won the Asia Research Award (Society of Chemical Engineers, Japan), Outstanding Research Award (Ministry of Science and Technology, 2006, 2014), BEST Biochemical Engineering Achievement Award, Wu Ta-You Memorial Award (NSC), Outstanding Academia-Industry Research Award and Outstanding Young Investigator Award in Taiwan. He is inducted as a fellow of American Institute for Medical and Biological Engineering (AIMBE), and is elected the Member of the Tissue Engineering International & Regenerative Medicine Society-Asia Pacific (TERMIS-AP) Council and the Vice President of Biotechnology and Biochemical Engineering Society of Taiwan. He is the Program Chair of the TERMIS-AP meeting, 2016. He also sits on the editorial board of 10 international journals and currently serves as the associated editor of Current Gene Therapy and deputy editor of Journal of Taiwan Institute of Chemical Engineers.