The type I interferon-mediated immune response triggered by gRNA is associated with DDX58. (A) Genotypes of DDX58 knockout (KO) HeLa cell lines. DDX58 KO clones #1 and #2 have homozygous out-of-frame mutations at the DDX58 locus. The target sequence complementary to the sgRNA used for mutagenesis is underlined and the PAM sequence is shown in red. (B–D) Relative IFNB1 (B), DDX58 (C), and OAS2 (D) mRNA levels in wild-type (WT) and DDX58 KO cell lines at 24 h after transfection. Error bars, SEM; n = 3. (E) WT and DDX58 KO cell viabilities after RNP transfection were determined by WST-1 assays. Error bars, SEM; n = 3. Statistical significances among the different measurements were calculated using one-way ANOVA. (**) P < 0.01, (****) P < 0.0001.
