Fig. 1

The performance of the CRISPR-Un1Cas12f1 systems in human cells. a Schematic of the editing ability analysis of the engineered CRISPR-Un1Cas12f1 systems, which contain two nucleases (also see Additional file 1: Fig. S1a) combined with three engineered sgRNA, ge3.0, ge4.0, and ge4.1. b Tag-seq-based comparative analysis of wild-type Un1Cas12f1 (WT), and Un1Cas12f1-V3.1 (V3.1, a reported high-active variant) combined with the engineered sgRNAs (ge3.0, ge4.0, and ge4.1) targeted to twenty-one sites in HEK293T cells (also see Additional file 1: Fig. S4). The sgRNA reference is shown on the top and the on-target and the off-target cleavages are displayed without or with mismatches to the sgRNA reference by color highlighting. Sequencing read counts are shown to the right of each site. c The characteristics of the Un1Cas12f1 induced double-strand-breaks (DSBs). Each point was calculated as the ratio of the read count at each break site to the total break read counts at this sgRNA, and then pooled within the twenty-one sgRNAs. Read counts were obtained from Tag-seq (Additional file 1: Fig. S5). x-axis shows the location of the sgRNA. The red dotted line indicates the reported in vitro DSB sites, while the blue dotted line indicates another potential breakpoint in vivo. d Normalization of on-target activity of the various CRISPR-Un1Cas12f1 systems to Un1Cas12f1-V3.1 + ge4.1 combination, value = (other system on-target reads)/(V3.1 + ge4.1 on-target reads). Grey points, sites without detectable editing. e Total number of off-target sites detected within the twenty-one sgRNAs. f Specificity Index assessment (value was calculated by the ratio of total on-target reads to the on-target reads plus the off-target reads within the twenty-one sites)