Figure 4
From: Deep mutational scanning of S. pyogenes Cas9 reveals important functional domains
Activity of negative and positive selection enriched mutants. Mutations which were enriched following both positive and negative selection were expressed in mammalian cells, targeting a chromosomally integrated copy of GFP. Differences in on-target activity were assessed with the site 1 (S1) gRNA (n = 3, error bars indicate S.E.M., one-way ANOVA F24,50 = 6.34, P = 2.1 × 10−8, followed by post hoc linear contrasts). We tested the ability of mutants to cleave a doubly mismatched S1 target (n = 3, error bars indicate S.E.M., one-way ANOVA F24,50 = 9.16, P = 3.4 × 10−11, followed by post hoc linear contrasts) and a singly mismatched target (n = 3, error bars indicate S.E.M., one-way ANOVA F24,50 = 11.37, P = 5.7 × 10−13, followed by post hoc linear contrasts, P = * < 0.05, ** < 0.01, *** < 0.001). Bars above each graph indicate the positions of mismatches with red colored boxes.
