Figure 4.

Gene editing activities of miniAGBEs at three target sites in PFFs and porcine embryos. (A) Comparison of A-to-G, C-to-G and C-to-T editing frequencies at 3 endogenous porcine genomic loci by miniAGBE-2, miniAGBE-3, and miniAGBE-4, respectively (n = 3) in PFFs (The indistinctive conversion of C-to-A are not shown). (B) Comparison of base editing products distribution (top) and indel frequencies (below) among edited porcine genomic loci in PFFs treated with miniAGBE-2, miniAGBE-3 and miniAGBE-4 and the corresponding sgRNA, or in control groups. Values and error bars indicate the mean ± s.d. of three independent replicates. Editing frequencies reflect sequencing reads that contain base editing only and do not contain indels among all treated cells, with puromycin selecting. PFFs electro-transfected with resuspension buffer only served as WT control. ns, no significant difference (P> 0.05). (C) Summary of porcine embryo development with miniAGBE-4. Embryo injected with sterile water served as WT control. ns, no significant difference (P> 0.05). (D) Scatter plots shows base editing frequencies in porcine embryos by co-delivering miniAGBE-4 mRNA (150 ng/μl) with each three sgRNAs (50 ng/μl) in (A) via micro-injection. Each dot indicates an individual embryo. Data are presented as mean ± s.d., and the bold lines are represented the mean of base editing frequencies.