Fig. 5: Mouse mitochondrial ND5 nonsense mutation generated via cytidine-deaminase-mediated base editing.

a The DdCBE target for generating the m.C12336T nonsense mutation and m.G12341A silent mutation. The m.C12336T (C₉) mutation creates a Q199stop mutation in the ND5 protein, whereas m.G12341A (C₁₄) causes a silent Q200Q mutation. Translation triplets are underlined and possible editing loci are shown in red. b The efficiency of the cytosine-to-thymine base editing that creates a nonsense mutation in NIH3T3 cells. The annotations indicate the combination of DdCBE pairs that were co-transfected into cells. Dark and light gray bars represent the frequency of m.C12336T (C₉) and m.G12341A (C₁₄) mutations, respectively. Error bars represent s.e.m. for n = 3 biologically independent samples (n.s. not significant, ^*p < 0.05, and ^**p < 0.01 using Student’s two-tailed t test). P values of left-G1333-N + right-G1333-C, left-G1333-C + right-G1333-N, left-G1397-N + right-G1397-C, and left-G1397-C + right-G1397-N for C₉ mutation are 0.0065, 0.1143, 0.0266, and 0.0037, and for C₁₄ mutation are 0.0077, 0.0144, 0.0406, and 0.0214, respectively. c Base editing efficiency in mouse blastocysts. The sequencing data were obtained from blastocysts that developed after zygotes were microinjected with mRNA encoding left-G1333-N and right-G1333-C DdCBE. Dark and light gray bars represent the frequency of the C₉ and C₁₄ mutations, respectively. d Alignment of mutant sequences from newborn pups. Edited bases are shown in red, and the column on the right indicates the editing frequencies in the mutant mitochondrial genome. e Sanger sequencing chromatograms from non-edited and edited mice. The red arrows indicates the substituted nucleotides. Source data are provided in the Source data file.