Details for the general specificity of SpCas9 variants.
Figure. Specificity of SpCas9 variants against the mismatches between the guide sequence of sgRNA and target sequence. (
a) A heat map showing specificity of SpCas9 variants against single-base mismatches between the guide RNA and target sequence. The specificity was calculated as 1 - (indel frequencies at mismatched target sequences divided by those at perfectly matched targets). The general specificity represents the average of specificity at all 20 positions. n = 21 - 24 target sequences for each position. (
b) Effects of mismatch types on activities of SpCas9 variants. Relative indel frequencies at targets with different types of one-base mismatches are shown. The boxes represent the 25th, 50th, and 75th percentiles; whiskers show 10th and 90th percentiles. n = 76 - 79, 76 - 79, and 304 - 306 target sequences for wobble transition, non-wobble transition, and transversion mismatches, respectively. (
c, d) Effect of the number of mismatched bases on the activities of SpCas9 variants. Relative indel frequencies at two- (c) and three- (d) base mismatched target sequences are shown. n = 138 - 139 (c) and 133 – 135 (d) target sequences. The boxes represent the 25th, 50th, and 75th percentiles; whiskers show 10th and 90th percentiles.
