Figure 3.
Massive off-target activities of three URR-targeted ZFNs
(A) Flowcharts of ZFN off-target identification algorithm. (B) Summary of log10-transformed normalized on-target reads, off-target reads, and off-target counts for three ZFNs at 72 and 96 h. (C) Schematic of on-targets and top 10 off-targets of three ZFNs at 72 and 96 h. The on-target was at the top line followed by off-targets with mismatches highlighted in color. The specific cleavage positions and normalized GUIDE-seq read counts are shown at the right of each site. (D) Correlation of normalized GUIDE-seq reads between 72 and 96 h in common off-target sites for three ZFNs. Spearman correlation coefficients are shown. (E) Summary of off-target counts of original and extended modes for three ZFNs. (F) Correlation between MIP indel rates and normalized GUIDE-seq reads for Wolfe-165 common off-targets at 72 and 96 h. The Spearman correlation coefficients for original and extended mode are shown separately. (G) WebLogo of ZFN Wolfe-161 off-targets. The contents form left to right of top panel are (1) the weblogs from off-targets sequences corresponding to left ZFP, (2) the distribution of spacer lengths from off-targets, and (3) the weblogs from off-target sequences corresponding to the right ZFP. The contents from left to right of bottom panel are (1) the distribution of off-target mismatches to intended sequences of the left ZFP and each zinc finger, (2) the distribution of off-target mismatches to intended sequences of Wolfe-161 ZFN, and (3) the distribution of off-target mismatches to intended sequences of right ZFP and each zinc finger.