Skip to main content
. 2015 Sep 10;5:13878. doi: 10.1038/srep13878

Figure 1. Evaluation of sgRNA:Cas9-mediated modifications of MSTN and FGF5 in goat fibroblasts.

Figure 1

(a) Schematic diagram of MSTN and FGF5 partial protein coding region and the targeting loci of sgRNA:Cas9. sgRNAs targeting sites are presented in red. PAM sequences are highlighted in green and underlined. (b) Left panel, PCR products of the targeted exon 2 of MSTN from goat fibroblasts transfected with Cas9 and MSTN sgRNA1. Right panel, detection of sgRNA:Cas9-mediated on-target cleavage of MSTN by T7E1 cleavage assay. PCR products from left panel were subjected to T7E1 cleavage assay. M, marker; WT, wild type PCR product from fibroblasts that have not treated with CRISPR/Cas9. (c) Left panel, PCR products of the targeted exon 3 of MSTN from goat fibroblasts transfected with Cas9 and MSTN sgRNA2. Right panel, detection of sgRNA:Cas9-mediated on-target cleavage of MSTN by T7E1 cleavage assay. PCR products from left panel were subjected to T7E1 cleavage assay. (d) Sequences of modified MSTN alleles. Target sequences complementary to MSTN sgRNAs are in red text; the mutations are blue, lower case; insertions (+), deletions (−) or mutation (m) are shown to the right of each allele. (e) PCR products of the targeted exon 1 of FGF5 from goat fibroblasts transfected with Cas9 and FGF5 sgRNA1, FGF5 sgRNA2 and FGF5 sgRNA1 & 2, respectively. (f) Detection of sgRNA:Cas9-mediated on-target cleavage of FGF5 by T7E1 cleavage assay. PCR products from (e) were subjected to T7E1 cleavage assay. (g)Sequences of modified FGF5 alleles. Target sequences complementary to FGF5 sgRNAs are in red text; the mutations are blue, lower case; insertions (+), deletions (−), mutation (m) or turnover (t) i shown to the right of each allele.