CRISPR-Cas9 genome editing activity in transfected ovine cells.

<p><b>A)</b> Exon 1 of the ovine <i>MSTN</i> gene. The sequence recognized by sgRNA is in italic and bold, from nt 110–130, the PAM sequence TGG is underlined and the rest of exonic sequences in smaller font. <b>B)</b> Ovine cells were transfected with pX330-cas9-<i>MSTN</i> plasmid co-expressing Cas9 and sgRNA and three days later cellular DNA was isolated and subjected to T7EI assay for detecting mutation at the targeted <i>MSTN</i> exon 1. The PCR amplifies a band of 634 bp and uncleaved (asterisk) and cleaved (arrows, 255 bp and 379 bp) products are indicated. Numbers below each lane indicate the percentage of targeted indels. NT: non-transduced cells, T: transfected cells, L: 1Kb DNA. The experiment is representative of two replicates performed with similar results.