title: Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects.
abstract: Bacterial RNA-directed Cas9 endonuclease is a versatile tool for site-specific genome modification in eukaryotes. Co-microinjection of mouse embryos with Cas9 mRNA and single guide RNAs induces on-target and off-target mutations that are transmissible to offspring. However, Cas9 nickase can be used to efficiently mutate genes without detectable damage at known off-target sites. This method is applicable for genome editing of any model organism and minimizes confounding problems of off-target mutations.
authors.affiliation: 1] Ministry of Education Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, Nanjing, China. [2].
authors.forename: Bin
authors.initials: B
authors.lastname: Shen
authors.affiliation: 1] Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK. [2].
authors.forename: Wensheng
authors.initials: W
authors.lastname: Zhang
authors.affiliation: 1] Ministry of Education Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, Nanjing, China. [2].
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Ministry of Education Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, Nanjing, China.
authors.forename: Jiankui
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Ministry of Education Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, Nanjing, China.
authors.forename: Jianying
authors.initials: J
authors.lastname: Wang
authors.affiliation: Ministry of Education Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, Nanjing, China.
authors.forename: Li
authors.initials: L
authors.lastname: Chen
authors.affiliation: Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Wang
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK.
authors.forename: Alex
authors.initials: A
authors.lastname: Hodgkins
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK.
authors.forename: Vivek
authors.initials: V
authors.lastname: Iyer
authors.affiliation: Ministry of Education Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, Nanjing, China.
authors.forename: Xingxu
authors.initials: X
authors.lastname: Huang
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK.
authors.forename: William C
authors.initials: WC
authors.lastname: Skarnes
mesh_terms: D000818:Animals; D001483:Base Sequence; D064113:CRISPR-Cas Systems; D003850:Deoxyribonuclease I / Q000235:genetics / Q000378:metabolism*; D004622:Embryo, Mammalian; D004720:Endonucleases / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D016680:Genome, Bacterial; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D009154:Mutation; D012329:RNA, Bacterial
chemical_list: D012329:RNA, Bacterial; D004720:Endonucleases; D003850:Deoxyribonuclease I