Fig. 3

Specificity comparison of the DNA targeting CRISPR systems in human cells. a Schematic of the editing specificity analysis by Tag-seq among the DNA targeting CRISPR editors (CRISPR-AsCas12a, CRISPR-LbCas12a, CRISPR-Un1Cas12f1, CRISPR-AsCas12f1, and CRISPR-SpCas9 systems). b Tag-seq-based comparative analysis of DNA targeting CRISPR systems with twenty-one sgRNAs (also see Additional file 1: Figs. S9–S12). The targeted sites for Cas12 and Cas9 share a common spacer sequence as shown at the top. As for Cas12a/Cas12f1, the sgRNA reference is the full sequence. As for SpCas9, the sgRNA sequence begins after the TTTR and ends with its NGG PAM. c The characteristics of the DNA targeting CRISPR systems induced DSBs revealed by Tag-seq. x-axis showing the location of the sgRNA. The red dotted line indicates the expected DSB sites, while the blue dotted line indicates the new potential breakpoint. d Total number of off-target sites detected with the twenty-one sgRNAs. N.E., no editing detected. e 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)