Fig. 1.

Single-gene base mutation of GGTA1, B4galNT2, CMAH in porcine blastocysts, and PFFs with BE4-Gam system. a The target sequences at the GGTA1, B4galNT2, and CMAH locus. The PAM sequence, sgRNA-target sequences, and substituted bases are shown in blue, black, and red, respectively. Representative sequencing chromatograms at the GGTA1 (b), B4galNT2 (c), and CMAH (d) targets of WT and edited porcine blastocysts. Target sequence (black), PAM region (blue), target sites (red), and mutant amino acid (bold and italic). Representative sequencing chromatograms at the GGTA1 (e), B4galNT2 (f), and CMAH (g) targets of WT and edited PFFs. Target amino acid are indicated by red box. The efficiency of mutation was predicted with online tool EditR (https://moriaritylab.shinyapps.io/editr-master). h The blastocysts of un-injected blastocysts and the blastocysts which injected single sgRNAs plus BE4-Gam. The blastocysts was stained with DAPI, and the nucleus in the blastocyst was shown in blue dots