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. 2016 Aug 26;6:32271. doi: 10.1038/srep32271

Figure 3. Evaluation of sgRNA:Cas9-mediated genetic modifications in lambs.

Figure 3

(a) PCR products of the targeted region of the MSTN, ASIP, and BCO2 genes of founder sheep co-microinjected with a mixture of Cas9 mRNA and sgRNAs. D2000 DNA Marker was used as a marker reference. (b) Detection of sgRNA:Cas9-mediated on-target cleavage of the MSTN, ASIP and BCO2 genes by using a T7EI cleavage assay. All PCR products from (a) were subjected to the T7EI cleavage assay. (c) Sequencing results of the modified MSTN, ASIP and BCO2 loci that were detected in the founder animals. Target sequences complementary to sgRNAs of targeted genes are in red text, while the PAM sequences are marked in green. The mutations are marked in blue, dashlines indicates deletions, and lowercase indicates insertions or replacements. Insertions (+), deletions (−), mutations (m) is shown to the right of each allele. The genotypes are shown to the right with the rates of total clones for TA-sequencing.