Highly efficient RNA-guided base editing in mouse embryos

• Published in: Nature biotechnology Vol. 35; no. 5; pp. 435 - 437
• Main Authors: Kim, Kyoungmi, Ryu, Seuk-Min, Kim, Sang-Tae, Baek, Gayoung, Kim, Daesik, Lim, Kayeong, Chung, Eugene, Kim, Sunghyun, Kim, Jin-Soo
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.05.2017; Nature Publishing Group
• Subjects: 45/41; 631/1647/1513/1967/3196; 631/1647/334/2045; Agriculture; Animals; Base Pairing - genetics; Bioinformatics; Biomedical Engineering/Biotechnology; Biomedicine; Biotechnology; brief-communication; Clustered Regularly Interspaced Short Palindromic Repeats - genetics; CRISPR-Associated Proteins - genetics; Cytidine Deaminase - genetics; Deoxyribonucleic acid; DNA; DNA repair; Embryo; Embryo, Mammalian - embryology; Embryo, Mammalian - physiology; Gene Editing - methods; Genetic aspects; Genetic disorders; Genetic engineering; Genetic recombination; Life Sciences; Methods; Mice; Mice, Inbred C57BL; Mice, Inbred ICR; Mutagenesis, Site-Directed - methods; Mutation; Point Mutation - genetics; Recombinant Fusion Proteins - genetics; RNA; RNA - genetics
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/nbt.3816

Mice with targeted point mutations are generated efficiently using Cas9–cytidine deaminase fusions. Base editors (BEs) composed of a cytidine deaminase fused to CRISPR–Cas9 convert cytidine to uridine, leading to single-base-pair substitutions in eukaryotic cells. We delivered BE mRNA or ribonucleoproteins targeting the Dmd or Tyr gene via electroporation or microinjection into mouse zygotes. F0 mice showed nonsense mutations with an efficiency of 44–57% and allelic frequencies of up to 100%, demonstrating an efficient method to generate mice with targeted point mutations.