文献类型：期刊文献

英文题名：CRISPR/Cpf1 facilitated large fragment deletion in Saccharomyces cerevisiae

作者：Li, Zhen-Hai[1];Liu, Min[1];Lyu, Xiao-Mei[2];Wang, Feng-Qing[1];Wei, Dong-Zhi[1]

机构：[1]East China Univ Sci & Technol, Newworld Inst Biotechnol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Nanyang Technol Univ, Sch Chem & Biomed Engn, Coll Engn, Singapore, Singapore

年份：2018

卷号：58

期号：12

起止页码：1100

外文期刊名：JOURNAL OF BASIC MICROBIOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000451862800010)】；

基金：National Natural Science Foundation of China, Grant number: 31500043

语种：英文

外文关键词：CRISPR/Cpf1; large fragment deletion; Saccharomyces cerevisiae

摘要：In this study, we focused on the applicability of CRISPR/Cpf1 in genome simplification of Saccharomyces cerevisiae and established a CRISPR/Cpf1 assisted method for rapid markerless large fragment deletion to facilitate laboratory evolution of geome of S. cerevisiae by rational genetic engineering. This method uses a Cpf1 expression plasmid and a crRNA array expression plasmid. The DNA fragment between two DSBs generated by CRISPR/Cpf1 can be cut off from the chromosome, along with re-ligation of the genomic endpoints of the DSBs. Using this method, the large DNA fragment of similar to 38 kb between the two genes of TRM10 and REX4 was successfully and rapidly deleted, which was verified by PCR and Sanger DNA Sequencing. This method is simple and rapid, and can be easily implemented for large fragment deletion in S. cerevisiae.