Mitochondrial DNA editing in mice with DddA-TALE fusion deaminases

• Published in: Nature communications Vol. 12; no. 1; p. 1190
• Main Authors: Lee, Hyunji, Lee, Seonghyun, Baek, Gayoung, Kim, Annie, Kang, Beum-Chang, Seo, Huiyun, Kim, Jin-Soo
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 19.02.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 13/109; 38/23; 631/1647/1511; 631/61/17/1511; 64/60; Animal models; Animals; Cytosine; Dehydration; Deoxyribonucleic acid; DNA; DNA, Mitochondrial - genetics; Editing; Electron transfer; Electron Transport; Embryos; Female; Gene Editing - methods; Genes, Mitochondrial - genetics; Humanities and Social Sciences; Male; Mice; Mice, Inbred C57BL; Mitochondria; Mitochondrial Diseases - genetics; Mitochondrial DNA; Mitochondrial Proteins - genetics; multidisciplinary; Mutation; NADH; NADH dehydrogenase; NADH Dehydrogenase - genetics; Nicotinamide adenine dinucleotide; NIH 3T3 Cells; Nonsense mutation; Science; Science (multidisciplinary); Stop codon; Toxins; Transcription; Transcription Activator-Like Effectors - genetics; Ubiquinone; Uracil
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-21464-1

DddA-derived cytosine base editors (DdCBEs), composed of the split interbacterial toxin DddA tox , transcription activator-like effector (TALE), and uracil glycosylase inhibitor (UGI), enable targeted C-to-T base conversions in mitochondrial DNA (mtDNA). Here, we demonstrate highly efficient mtDNA editing in mouse embryos using custom-designed DdCBEs. We target the mitochondrial gene, MT-ND5 ( ND5 ), which encodes a subunit of NADH dehydrogenase that catalyzes NADH dehydration and electron transfer to ubiquinone, to obtain several mtDNA mutations, including m.G12918A associated with human mitochondrial diseases and m.C12336T that incorporates a premature stop codon, creating mitochondrial disease models in mice and demonstrating a potential for the treatment of mitochondrial disorders. Split DddA-derived base editors fused to TALEs enable mitochondrial DNA editing. Here the authors demonstrate their use in mouse embryos with germline transmission.