Figure 8
From: Deep mutational scanning of S. pyogenes Cas9 reveals important functional domains
Activity of positive selection enriched mutant combinations. (a) Combinations of mutations that had retained activity in mammalian cells as single mutants were compared to WT SpCas9’s ability to cleave with a gRNA targeting GFP site 4 (S4) (n = 3, error bars indicate S.E.M., one-way ANOVA F25,52 = 5.07, P = 4.0 × 10−11, followed by post hoc linear contrasts). (b) Top performing combinations were targeted against chromosomally integrated GFP with a total of eight gRNAs and their activity was normalized between WT SpCas9 with each gRNA and a non-targeting gRNA (one-way ANOVA F6,161 = 3.65, P = 0.0020, followed by post hoc linear contrasts, P = * < 0.05, ** < 0.01, *** < 0.001).
