Fig. 2

Targeted mutagenesis in mice using a prime editor 3 with proximal dsgRNA. a A schematic of the mutagenesis design at exon 4 in Igf2. b The genotypes of two mice harboring G to C conversion and TA insertion mutations induced by PE3 with proximal dsgRNA +7 in Igf2. c Sanger-sequencing chromatograms of the two mice harboring the desired mutations. The green arrow denotes the G to C conversion for modification of the PAM sequence, while the red arrow denotes the TA insertion for the stop codon. d Whole-genome sequencing analysis of the Igf2 wild-type (WT) and mutant #1 mice. The potential off-target (OT) sites of pegRNA, nsgRNA, and dsgRNA were obtained using Cas-OFFinder. e Sequence alignment and Sanger-sequencing chromatogram of potential OT sites. f An image of the offspring after mating an Igf2 p+/m− male (F1) with an Igf2 p+/m+ female: wild-type (left) and Igf2 mutant (right). g Bodyweight 2 weeks after birth. WT, wild-type; MUT, mutant; p, paternal allele; m, maternal allele. Error bars show the mean ± standard deviation (s.d.)