Fig. 1
From: Efficient generation of mouse models of human diseases via ABE- and BE-mediated base editing
ABE-mediated efficient A-to-G conversion at Ar and Hoxd13 loci in mouse embryos. a GFP-to-BFP conversion as a reporter for ABE-mediated base editing. b Analysis of base editing by FACS (left) and Sanger sequencing (right). Scramble: Scrambled sgRNA as negative control; sgRNA: sgRNA targeting GFP; PC: BFP expression plasmid only as positive control. Data were analyzed by Student’s t-test (***p < 0.001) and shown as mean ± s.e.m. (n = 3 from three independent experiments). c ABE-mediated base editing in vivo. ABE mRNA and sgRNA were co-injected into one-cell embryos, and the editing efficiency examined at blastocyst stage. d Efficiency of A-to-G substitution in mouse embryos. TA clones of PCR amplicons from the target regions in Ar (for sgAr-1, sgAr-15) and Hoxd13 (sgHoxd13) were analyzed by Sanger sequencing. Each dot indicates one individual mouse. At least 10 TA clones were analyzed for each sample. e The editing frequencies of individual A-to-G conversion of samples described in c were analyzed. A3, A7, C5, and T11 indicate edited positions of the protospacer for sgAr-1; A6, A8, and A9 indicate edited positions of the protospacer for sgHoxd13. f Representative alignments of modified sequences from embryos after microinjection of ABE mRNA and sgRNAs into one-cell embryos. The PAM sequences and substitutions are highlighted in red and blue, respectively; the target codons are underlined; N/N represents positive colonies out of the total sequenced
