Fig. 8: Target-matched nucleases show high efficiency without any genome-wide off-target for targets tested in this study regardless their ranking.

a Summary of targets edited by IFNs, examined in this study with GUIDE-seq and NGS. For 10 target sites, including those challenging targets where previous attempts with IFNs had failed, we were able to perform editing without any off-target detected by GUIDE-seq and further confirmed by NGS on the top three site. For the highest ranked target, CCR5 site 11, NGS still identified residual off-target activity even with the highest ranked B-HeF, indicating that development of an even higher fidelity IFN would be beneficial for accessing the highest cleavability rank. The colors of the squares indicate the percentage value of the on-target genome modification normalized to WT (measured by NGS). Colored circles indicate the summarized GUIDE-seq and NGS results; green circle indicates when both NGS and GUIDE-seq showed no off-targets, red circle indicates off-target editing detected either by GUIDE-seq or NGS, light green circle indicates when no off-target was found but it was only tested by NGS and gray circle indicate no data. Off-target editing data of these targets (GUIDE-seq experiments) from the literature are summarized in Supplementary Data file 6: Data from other studies. The ranking of the targets is weakly related to either b, the number of their predicted off-target sites, or c, the detected off-target sites using WT SpCas9 (for details see Supplementary Table 2). a-c Data are related to Figs. 6, 7, 9, Supplementary Figs. 9–11.