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. 2015 Nov 10;10(11):e0142755.
doi: 10.1371/journal.pone.0142755. eCollection 2015.

Simple Genome Editing of Rodent Intact Embryos by Electroporation

Takehito Kaneko  1 , Tomoji Mashimo  2
Affiliations

Simple Genome Editing of Rodent Intact Embryos by Electroporation

Takehito Kaneko et al. PLoS One. .

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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Conflict of interest statement

Competing Interests: Kyoto University and NEPA GENE Co. Ltd. have obtained a patent for this method (patent no. 5774657). T.K. and T.M. are listed as inventors of the patent and will receive part of the revenue that it may generate. This does not alter the authors' adherence to PLOS ONE policies on sharing data and materials.

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