Fig. 3

Recovery of editing efficiency of high-fidelity Cas9 variants using HH ribozyme-linked sgRNAs. a A schematic of a self-processing ribozyme fused sgRNA. The pre-sgRNA contains a HH ribozyme at its 5’-end. The pre-sgRNA undergoes self-cleavage to release a mature sgRNA. The red arrow indicates the self-cleavage position. b HH ribozyme-fused sgRNAs with a matched 5’ nucleotide (HH-X₂₀) or a mismatched guanosine (HH-gX₁₉) were tested in combination with Cas9-WT and high-fidelity Cas9 variants at six target sites in HeLa cells. Indel frequencies were measured using targeted deep sequencing. The PAM is shown in blue. Error bars, s.e.m. of three biological replicates. Statistical significances were calculated by unpaired t-test. * P < 0.05, ** P < 0.01. c Mean indel frequencies ± s.e.m. at the six target sites in HeLa cells. Statistical significances were calculated by paired t-test. ** P < 0.01, *** P < 0.001