Fig. 4
BhCas12b v4 is a highly specific nuclease. a Comparison of Cas9, Cas12b, and Cas12a indel activity in 293T cells at nine target sites (except for Cas12a, which was only tested at the three TTTV PAM sites) selected for Guide-Seq analysis. Error bars represent s.d. from n = 4 replicates. b Guide-Seq analysis showing the number and relative proportion of detected cleavage sites for each nuclease. Off-targets are shown as light gray wedges, while the on-target site is highlighted in purple (for SpCas9), dark blue (for BhCas12b v4), or light blue (for AsCas12a) with the fraction of on-target reads shown below. Off-targets were only detected with SpCas9. n.t., not tested. See Supplementary Fig. 6 for full analysis. c BhCas12b indel activity in 293T cells when mismatches are present between the guide sgRNA and target DNA. Mismatches were inserted in the sgRNA to match the target strand (i.e., C to G, A to T). Error bars represent s.d. from n = 4 replicates. Source data are provided as a Source Data file
