CRISPR/Cas9-mediated modification of the BLG locus in fibroblasts.
figureposted on 2017-10-10, 17:38 authored by Wenjun Zhou, Yongjie Wan, Rihong Guo, Mingtian Deng, Kaiping Deng, Zhen Wang, Yanli Zhang, Feng Wang
(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.