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• W3001434723 abstract "Genome edited animals can now be easily produced using the clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated protein 9 (Cas9) system. Traditionally, these animals have been produced by the introduction of endonucleases into pronuclear-stage embryos. Recently, a novel electroporation method, the “Technique for Animal Knockout system by Electroporation (TAKE),” has been established as a simple and highly efficient tool to introduce endonucleases into embryos instead of methods such as microinjection. Use of frozen-warmed pronuclear-stage embryos in this method has further contributed to efficient production of genome edited animals. However, early developmental stage embryos, including pronuclear-stage embryos, especially those of rats, sometimes show low resistance to physical damage by vitrification and introduction of endonucleases during microinjection. In this study, we propose an ethanol-free, slow-freezing method to reduce physical damage to pronuclear-stage embryos followed by the TAKE method. All mouse and rat frozen embryos were survived after electroporation, and 18% and 100% of offspring were edited target gene, respectively. The resulting protocol is an efficient method for producing genome edited animals." @default.
• W3001434723 created "2020-01-30" @default.
• W3001434723 creator A5048938769 @default.
• W3001434723 creator A5056971008 @default.
• W3001434723 date "2020-02-01" @default.
• W3001434723 modified "2023-10-15" @default.
• W3001434723 title "Genome editing of rodents by electroporation of CRISPR/Cas9 into frozen-warmed pronuclear-stage embryos" @default.
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• W3001434723 doi "https://doi.org/10.1016/j.cryobiol.2020.01.016" @default.
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