Figure 3: Genome-wide target specificities of CjCas9 nucleases examined using Digenome-seq.
From: In vivo genome editing with a small Cas9 orthologue derived from Campylobacter jejuni
Human or mouse genomic DNA isolated from HeLa cells or NIH 3T3 cells (grey), respectively, was digested in vitro by Cas9 and its sgRNA targeted to the human AAVS1 locus (a,b,f) and the mouse Rosa26 (c), Vegfa (c,d) and Hif1a loci (d) and subjected to whole-genome sequencing. Circos plots show genome-wide DNA cleavage scores across the human or mouse genome. Red arrows indicate on-target sites. N indicates the number of in vitro cleavage sites identified by Digenome-seq. (a,b) Sequence logos were obtained by comparing DNA sequences at in vitro cleavage sites with each other. (b) Indel frequencies at the AAVS1-TS8 site measured using targeted deep sequencing. CjCas9 (orange) or SpCas9 (blue) targeted to the AAVS1-TS8 site was transfected into human HEK293 cells. Error bars indicate s.e.m. (n=3). (e) Indel frequencies at three AAVS1 sites targeted by CjCas9 (orange) and SaCas9 (violet).
