Fig. 2.

The generation of Ar and Hoxd13 mutant mouse models by ABE. a The editing efficiencies were detected by Sanger sequencing of TA clones of mouse pups for sgAr-1, sgAr-15, and Hoxd13, respectively. Each dot indicates one individual mouse. At least ten TA clones were analyzed for each sample. Blue dots represent male mice and red dots represent female mice. Asterisks represent the mice with defects. b The editing efficiencies of different positions from individual samples described in a were analyzed. Left: A₃, A₇, and C₅ indicate the edited positions of the protospacer for sgAr-1; A₆, A₈, and A₉ indicate edited positions of the protospacer for sgHoxd13. Blue dots represent male mice and red dots represent female mice. Right: Editing frequencies in As of each mice with defects for sgHoxd13 and sgAr-1 are displayed in heatmap. Blue rectangles represent male mice and red rectangles represent female. Editing frequencies increase with the increase of color shades. c Representative alignments of modified sequences from pups 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. d Characterization of the targeted modifications by deep sequencing. The frequencies were calculated from three (n = 3) replicates. e Sex reversal in founder mouse. Left: a 5-week-old mouse with female genitalia (green arrowhead) and nipples (red arrowheads); middle: Wt male with normal male genital (blue arrowhead); right: founder A021 with internal genitalia of male (orange arrowhead) and smaller testes and founder A063 with smaller testes. f Syndactyly phenotypes of founder mouse. Digits of mutant mouse are fused (purple arrowhead)