Figure 4: AAV-mediated mutagenesis in vitro and in vivo.
From: In vivo genome editing with a small Cas9 orthologue derived from Campylobacter jejuni
(a) AAV vector encoding CjCas9 and its sgRNA. (b) Indel frequencies at the Rosa26 target site in mouse C2C12 myotubes infected with AAV-CjCas9. (c) Indel frequencies at the Rosa26 target site in TA muscles of C57BL6J mice injected with AAV-CjCas9 measured at 2, 4, 8 and 32 weeks after injection. One-way ANOVA and Tukey’s post hoc tests, **P<0.01, NS, not significant. See also Supplementary Fig. 3. (d) No off-target indels were detected at 20 homologous sites that differed from the on-target site by up to 4 nucleotides in the mouse genome. Genomic DNA isolated from AAV-CjCas9-injected TA muscles of C57BL6J mice mice at 32 weeks after injection was analysed by targeted deep sequencing. Mismatched nucleotides are shown in blue and PAM sequences in red. Red arrows indicate cleavage positions within the 20-bp target sequences. Error bars indicate s.e.m. (n=3).
