Figure 4.

GKCR method highly knocked out HPV18 E6 and E7 oncogenes in the HeLa cells

(A and B) Agarose gel electrophoresis (A) and Sanger sequencing (B) of PCR products to verify that the Cas9/gRNA cassette integrated into the targeted HPV18 E6 and E7 locus. (C) Fold change of forward and reverse integration of the corresponding Cas9/gRNA cassettes into the respective target sites, analyzed by qPCR. Data are presented as mean ± SD (n = 3 per group). ∗∗p < 0.01, ∗∗∗p < 0.001; analyzed by two-tailed Students’ t test. (D) TIDE assays revealed that the GKCR method induced a significantly higher indel percentage than the control method at the HPV18 E6 and E7 locus. Data are presented as mean ± SD (n = 3 per group). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001; analyzed by one-way ANOVA and Tukey’s post hoc test.