Supplementary Figure 8: Evaluation of evoCas9 specificity by targeted deep-sequencing
From: A highly specific SpCas9 variant is identified by in vivo screening in yeast
(a) On-/off-target ratios for the nine common off-targets associated to the VEGFA2 locus calculated from the data in Fig. 3g and reported for wild-type SpCas9 and each different high-fidelity variant, as indicated in the graph. The bars indicate the interval between the values of the ratios obtained from the two replicate samples. Indel analysis of previously validated off-target sites relative to the EMX1 locus (b) and the VEGFA3 locus (c) performed by targeted deep-sequencing on genomic DNA of 293T cells expressing wild-type SpCas9, SpCas9-HF1, eSpCas9(1.1) or evoCas9 together with each specific sgRNA. Cells not expressing SpCas9 were sequenced to determine background indel levels. Two independent experiments were performed by mixing genomic DNA from three biological replicates before library preparation. The shaded circles represent the two measured editing percentages, while the solid circle indicates the mean.
