Figure 3.

Tolerance of Cas9 orthologs for truncated gRNAs. The efficiencies of RGN-mediated cleavage using truncated gRNA was assessed at human PRKDC (a) and CARD11 (b) loci. Genomic DNA was extracted 3 days upon transfection of HEK293T cells and extent of cleavage was monitored with T7EI assay. Untreated cells were used as a control (−). The lengths of the truncated gRNAs are indicated above each lane and mismatched 5′-G are highlighted with asterisks (*). Arrows indicate the positions of the expected cleavage products. The average frequencies of modified alleles are indicated at the bottom of each panel (n = 3). RGN, RNA-guided nuclease.