Fig. 2 | Nature Communications

Fig. 2

From: Pairwise library screen systematically interrogates Staphylococcus aureus Cas9 specificity in human cells

Fig. 2

SaCas9 sgRNA with shorter spacers are less tolerant of mismatches. a Average effect of sgRNA spacer length and mismatch position on SaCas9 single-mismatch tolerance. Mean ± 95% confidence interval is shown (n = 16,742 guide-target pairs). b Heatmap of relative SaCas9 cleavage efficiency for each possible RNA:DNA base pair, calculated for all single mismatch library members (n = 10,344 guide-target pairs). c Heatmap of relative SaCas9 double-mismatch tolerance across different spacer lengths and positions. Data is aggregated from 10 sgRNAs for which all possible single and double mismatches were tested (n = 28,707 guide-target pairs). d DNA base identity and position effect on double-mismatch tolerance for 20 nt spacer sgRNA (n = 6017 guide-target pairs) and e for 21 nt spacer sgRNA (n = 6729 guide-target pairs). All panels use data from Day 3 post-SaCas9 delivery

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