Figure 1.

Analysis of targeted KCNQ3 mutagenesis in DAT^Cremice. Analysis of FACS-sorted GFP+ (A) and GFP- (B) nuclei from mice co-injected with AAV1-FLEX-SaCas9-U6-sgKCNQ3 and AAV1-FLEX-EGFP-KASH into the VTA. (A₁) sgKcnq3 sequence with PAM underlined and SaCas9 cut site indicated by black arrow. Chromatogram results display multiple peaks beginning at the cut site. (A₂) Top 10 mutations at the cut site with the percent of total reads for which they occur on the left. Base changes: bolded. Insertions: underlined. Deletions: marked with a “-” (dash). (B) Chromatogram of GFP- nuclei sequencing displaying no evidence of mutations at the SaCas9 cut site. (C) Chromatogram of sgKcnq3 predicted off-target in Grm8 displaying no evidence of mutations at the SaCas9 cut site. (D) Frequency distribution of insertions and deletions in Kcnq3 from GFP+ nuclei. (E) Percent of wild-type, deletions, insertions, and base changes as percent of total reads for Kcnq3 in GFP+ (left) and GFP- nuclei (middle) and the predicted off-target Grm8 (right).