Fig. 7

Performance comparison of Cas12a variants. a The conserved sites of ultra variant in AsCas12a and LbCas12a, underlined with red. b The mutation sites of AsCas12a and LbCas12a variants. RKA=Plus, UKA=ultra+KA, UKK=ultra+KK, RU=high-active substitution E174R/D156R+ultra, RKAU=RKA+ultra, RKKU=RKK+ultra. c–e Tag-seq-based comparative analyses of wild-type AsCas12a (WT), and AsCas12a variants with twenty-eight sgRNAs targeting nineteen genes (see Additional file 2: Figure S7b). c Total number of off-target sites detected with the twenty-eight sgRNAs. d Specificity index (value was calculated by the ratio of total on-target reads to the on-target reads plus the off-target reads within the twenty-eight sites). e Normalization of on-target activity of AsCas12a variants to wild-type AsCas12a. f–h Tag-seq-based comparative analyses of wild-type LbCas12a (WT), and LbCas12a variants with fifteen sgRNAs targeting nine genes (see Additional file 2: Figure S8b). f Total number of off-target sites detected with the fifteen sgRNAs. g Specificity Index (value was calculated by the ratio of total on-target reads to the on-target reads plus the off-target reads within the fifteen sites). h Normalization of on-target activity of LbCas12a variants to wild-type LbCas12a