Endogenous gene disruption through CPP-mediated delivery of Cas9 protein and guide RNA. HEK293T cells were either sequentially or simultaneously treated with Cas9-m9R and sgRNA:9R three times. (A) Mutations detected by the T7E1 assay 3 d after the first treatment. The arrow indicates the expected position of DNA bands cleaved by T7E1. Mutations frequencies (indel [%]) were calculated from the band intensities. (B) DNA sequences of the CCR5 wild-type (WT) and mutant clones. The target sequence complementary to sgRNA is underlined. The protospacer adjacent motif is shown in bold. The cleavage site is indicated by an arrowhead. The column on the right indicates the number of inserted or deleted bases.
