Figure 1.

Chicken myostatin (MSTN) gene structure and targeted site design for nickase-mediated mutant induction. (A) Sequences of chicken MSTN exon1 and intron. The bold ATG indicates the start codon and the blue italic sequences present the forward and reverse primer for polymerase chain reaction amplification. The red sequences are the targeted sites of the CRISPR-associated protein 9 (Cas9)-D10A nickase. (B) The left and right target sequences (highlighted boxes) for the nickase. The target site has +7 bp offset and consequently, the targeted sites show 5′ overhanging 43 bp after the nickase-mediated DNA breaks. The red sequences are the targeted sites and the blue sequences are the complimentary sequences. The bold sequences indicate the protospacer adjacent motif (PAM). The arrow heads are the cleavage positions by the Cas-D10A nickase.