title: A CRISPR-Cas9 Assisted Non-Homologous End-Joining Strategy for One-step Engineering of Bacterial Genome.
abstract: Homologous recombination-mediated genome engineering has been broadly applied in prokaryotes with high efficiency and accuracy. However, this method is limited in realizing larger-scale genome editing with numerous genes or large DNA fragments because of the relatively complicated procedure for DNA editing template construction. Here, we describe a CRISPR-Cas9 assisted non-homologous end-joining (CA-NHEJ) strategy for the rapid and efficient inactivation of bacterial gene (s) in a homologous recombination-independent manner and without the use of selective marker. Our study show that CA-NHEJ can be used to delete large chromosomal DNA fragments in a single step that does not require homologous DNA template. It is thus a novel and powerful tool for bacterial genomes reducing and possesses the potential for accelerating the genome evolution.
authors.affiliation: State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.
authors.forename: Tianyuan
authors.initials: T
authors.lastname: Su
authors.affiliation: State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.
authors.forename: Fapeng
authors.initials: F
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.
authors.forename: Pengfei
authors.initials: P
authors.lastname: Gu
authors.affiliation: State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.
authors.forename: Haiying
authors.initials: H
authors.lastname: Jin
authors.affiliation: State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.
authors.forename: Yizhao
authors.initials: Y
authors.lastname: Chang
authors.affiliation: National Glycoengineering Center, Shandong University, Jinan, 250100, People's Republic of China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.
authors.forename: Quanfeng
authors.initials: Q
authors.lastname: Liang
authors.affiliation: State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China.|National Glycoengineering Center, Shandong University, Jinan, 250100, People's Republic of China.
authors.forename: Qingsheng
authors.initials: Q
authors.lastname: Qi
mesh_terms: D064113:CRISPR-Cas Systems / Q000235:genetics*; D002872:Chromosome Deletion; D002876:Chromosomes, Bacterial; D053903:DNA Breaks, Double-Stranded; D059766:DNA End-Joining Repair; D004926:Escherichia coli / Q000235:genetics; D018390:Gene Targeting; D005818:Genetic Engineering / Q000379:methods*; D016680:Genome, Bacterial; D007763:Lac Operon