title: Simple Genome Editing of Rodent Intact Embryos by Electroporation.
abstract: The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system is a powerful tool for genome editing in animals. Recently, new technology has been developed to genetically modify animals without using highly skilled techniques, such as pronuclear microinjection of endonucleases. Technique for animal knockout system by electroporation (TAKE) method is a simple and effective technology that produces knockout rats by introducing endonuclease mRNAs into intact embryos using electroporation. Using TAKE method and CRISPR/Cas system, the present study successfully produced knockout and knock-in mice and rats. The mice and rats derived from embryos electroporated with Cas9 mRNA, gRNA and single-stranded oligodeoxynucleotide (ssODN) comprised the edited targeted gene as a knockout (67% of mice and 88% of rats) or knock-in (both 33%). The TAKE method could be widely used as a powerful tool to produce genetically modified animals by genome editing.
authors.affiliation: Institute of Laboratory Animals, Graduate School of Medicine, Kyoto University, Kyoto, 606-8501, Japan.
authors.forename: Takehito
authors.initials: T
authors.lastname: Kaneko
authors.affiliation: Institute of Experimental Animal Sciences, Faculty of Medicine, Osaka University, Osaka, 565-0871, Japan.
authors.forename: Tomoji
authors.initials: T
authors.lastname: Mashimo
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D018274:Electroporation; D018390:Gene Targeting / Q000379:methods; D005818:Genetic Engineering / Q000379:methods; D016678:Genome; D051379:Mice; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems; D012333:RNA, Messenger