Fig. 2: Analysis of the PEAC-seq off-target sites.

a The visualization of PEAC-seq on-target and off-target sites. The ‘*’ represented a PEAC-seq site that was also called by the GUIDE-seq. The ‘**’ represented a PEAC-seq off-target (PEAC-seq-unique) that was identified by Amplicon-seq but not called by the GUIDE-seq. PEAC score: quantitative enrichment of the PEAC-seq tag at the edited sites; PEAC-ID: each identified site (on-target and off-target) by PEAC-seq were assigned a PEAC-ID, which was ordered by the PEAC score (descending order). b The number of reads from the shared PEAC-seq and GUIDE-seq sites is highly correlated. c Screenshots of PEAC-seq signal tracks from the IGV Genome Browser. One on-target site, one shared off-target site, and one PEAC-seq unique off-target site were presented. For each site, signals from both the PEAC-seq and the wild-type (WT, no Cas9-MMLV treatment) samples were included. For each sample, the first track represented signals from the amplicons of a forward primer and a downstream Tn5 primer; the second track represented signals from the amplicons of a reverse primer and an upstream Tn5 primer. The model on the right side showed the direction of the spacer and PAM of each case. d The shared off-targets (gray bars) tend to have less mismatches compared to the on-target site, while the PEAC-seq unique sites (orange bars) and the GUIDE-seq unique sites (blue bars) tend to have more mismatches. e. Mutation frequencies were plotted at each position alongside the gRNA and PAM sequences (from 5’ to 3’). From top to bottom are profiles of VEGFA TS1, TS2, and TS3. Source data are provided as a Source data file.