Fig. 3

Side-by-side comparison of SpCas9 variants programmed with perfectly matching sgRNAs reveal a target-selectivity ranking among the variants in the order of eSpCas9 > SpCas9-HF1 > HeFSpCas9. a EGFP disruption activities of the nucleases, calculated as described in “Methods”. Bars correspond to averages of n = 3 parallel samples; error bars represent the standard errors estimated by Gaussian error propagation of the component standard deviations associated with both EGFP and mCherry (transfection control) values. The target numbers in squares are the 16 targets examined in Fig. 4a. b Summary of the characteristics of distributions of data for on-target disruption activities of nuclease variants normalized to that of the WT SpCas9. Tukey-type notched boxplots by BoxPlotR: center lines show the medians; box limits indicate the 25th and 75th percentiles; whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles; notches indicate the 95% confidence intervals for medians; crosses represent sample means; data points are plotted as open circles. The sample points (24 in case of each variant) correspond to the targets present on Fig. 3a. Each pair of means is statistically different at the p < 0.05 level: SpCas9-HF1–eSpCas9 (<0.002), SpCas9-HF1–HeFSpCas9 (<0.001), HeFSpCas9–eSpCas9 (<0.001)