Fig. 1. Structure-guided mutagenesis increases on-target activity of SpCas9-HF1 with 21G-sgRNAs.

a X-ray crystallography derived structure of SpCas9-sgRNA-DNA complex in the conformation closest to the cleavage-competent state (PDB ID: 5f9r)⁴³. b Sequences of SpCas9-HF1 and the selected Blackjack-SpCas9-HF1 at the region affected, between residues L1004 and D1017; deletions (−) and insertions (green) are indicated. See also Supplementary Fig. 1. c, d Blackjack mutations increase on-target activities of increased fidelity variants with 21G-sgRNAs on different targets. Means are shown, error bars represent the standard deviation (s.d.) for n = 3 biologically independent samples (overlaid as white circles).