CRISPR/Cas9-mediated modification of the BLG locus in fibroblasts.

(A) Schematic diagram of sgRNA design for the goat BLG locus. Primers labeled as “BLG-T7-F” and “BLG-T7-R” were used for the T7E1 cleavage assay at the sg1, sg2, and sg3 target sites. (B) Targeting BLG loci using single sgRNA by electroporation. Top panel: PCR products of the target region of BLG from fibroblasts transfected with a single Cas9-sgRNA plasmid. Bottom panel: T7E1 assay of products shown in the top panel. M, marker; WT, wild-type cells without treatment with Cas9 plasmid. Red arrows indicate the expected cleaved products after T7E1 cleavage assay. (C) Sequencing results of sgRNAs targeting BLG.