Figure 1.

CRISPR/Cas9-mediated C3 gene targeting in PFFs. (a) Schematic diagram of sgRNA targeting the exon 26 of the C3 gene. sgRNA targeting sites are highlighted in red. PAM sequences are underlined and highlighted in blue. (b) T7E1 assay for CRISPR/Cas9-mediated cleavage at target site in PFFs. WT1 and WT2: PCR products of untransfected PFFs treated with T7E1; sgRNA1 and sgRNA2: PCR products of PFFs transfected with Cas9-sgRNA1 and Cas9-sgRNA2 treated with T7E1. (c) Biallelic mutation donor cell sequences for SCNT. The wild-type sequence is shown at the top of the target sequence. The sgRNA sequences are highlighted in red, PAM sequences in blue, and insertions in green; deletion (−); insertion (+).