文献类型：期刊文献

英文题名：Cpf1-assisted efficient genomic integration of in vivo assembled DNA parts in Saccharomyces cerevisiae

作者：Li, Zhen-Hai[1];Liu, Min[1];Wang, Feng-Qing[1];Wei, Dong-Zhi[1]

机构：[1]East China Univ Sci & Technol, Newworld Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2018

卷号：40

期号：8

起止页码：1253

外文期刊名：BIOTECHNOLOGY LETTERS

收录：;EI(收录号：20182205241276);WOS:【SCI-EXPANDED(收录号:WOS:000438090800011)】；

基金：This research was financially supported by the National Natural Science Foundation of China (No. 31500043). The authors sincerely thank Prof. Hongwei Yu (Zhejiang University) for providing plasmids of pESC-LEU-cen, pESC-URA, pMRI-34-crtE-tHMG1, pMRI-35-crtYB-crtI, pUMRI-15, and strain of S. cerevisiae BY4741.

语种：英文

外文关键词：CRISPR-Cpf1; Saccharomyces cerevisiae; Genomic integration

摘要：To test the applicability of Cpf1 from Francisella novivida in genomic integration of in vivo assembled DNA parts in Saccharomyces cerevisiae. An easy-to-use vector toolkit, containing a CEN6/ARS4 plasmid expressing Cpf1 from Francisella novivida (FnCpf1) and a 2 mu plasmid for crRNA or crRNA array expressing, was constructed for Cpf1-assisted genomic integration in S. cerevisiae. Our results showed that FnCpf1 allowed for targeted singleplex, doubleplex, and tripleplex genomic integration of in vivo assembled DNA parts with efficiencies of 95, 52, and 43%, respectively. CRISPR-Cpf1 system allows for efficient genomic integration of in vivo assembled DNA parts in S. cerevisiae, and thus provides an alternative CRISPR-Cas method for metabolic pathway engineering in addition to CRISPR-Cas9 system previously reported for yeast.