Simple Genome Editing of Rodent Intact Embryos by Electroporation |
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| Authors: | Takehito Kaneko Tomoji Mashimo |
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| Institution: | 1. Institute of Laboratory Animals, Graduate School of Medicine, Kyoto University, Kyoto, 606–8501, Japan.; 2. Institute of Experimental Animal Sciences, Faculty of Medicine, Osaka University, Osaka, 565–0871, Japan.; Ohio State University Comprehensive Cancer Center, UNITED STATES, |
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| 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. |
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