Adenine base editing in mouse embryos and an adult mouse model of Duchenne muscular dystrophy
 Authors and Affiliations
 Authors and Affiliations
Seuk-Min Ryu1,2,4, Taeyoung Koo1,4, Kyoungmi Kim1,3,4, Kayeong Lim1,2,4, Gayoung Baek1, Sang-Tae Kim1, Heon Seok Kim1,2, Da-eun Kim1,2, Hyunji Lee1, Eugene Chung1,2 & Jin-Soo Kim1,2,*
1Center for Genome Engineering, Institute for Basic Science, Seoul, Republic of Korea. 2Department of Chemistry, Seoul National University, Seoul, Republic of Korea. 3Department of Biomedical Sciences and Department of Physiology, Korea University College of Medicine, Seoul, Republic of Korea. 4These authors contributed equally to this work.
*Correspondence to Jin-Soo Kim
Abstract denine base editors (ABEs) composed of an engineered adenine deaminase and the Streptococcus pyogenes Cas9 nickase enable adenine-to-guanine (A-to-G) single-nucleotide substitutions in a guide RNA (gRNA)-dependent manner. Here we demonstrate application of this technology in mouse embryos and adult mice. We also show that long gRNAs enable adenine editing at positions one or two bases upstream of the window that is accessible with standard single guide RNAs (sgRNAs). We introduced the Himalayan point mutation in the Tyr gene by microinjecting ABE mRNA and an extended gRNA into mouse embryos, obtaining Tyr mutant mice with an albino phenotype. Furthermore, we delivered the split ABE gene, using trans-splicing adeno-associated viral vectors, to muscle cells in a mouse model of Duchenne muscular dystrophy to correct a nonsense mutation in the Dmd gene, demonstrating the therapeutic potential of base editing in adult animals.
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