Fig. 3

Targeted CRISPR/Cas9 inhibition of EBV LMP1 and LMP2A genes in CNE-2 cells. a: Schematic depicting the target sites CRISPR/Cas9 editing in LMP2A (upper) and latent membrane protein 1 (LMP1) (lower) genes of the EBV genome. The target sites of gRNAs and binding sites of specific primers (LMP1-F, LMP1-R, LMP2A-F, and LMP2A-R) are indicated with arrows. b: Green fluorescence intensity in CNE-2 cells transfected with the LMP1-CRISPR/Cas9 or LMP2A-CRISPR/Cas9 plasmid for 6 and 24 h. c and d: Editing effect of CRISPR/Cas9. CNE-2 cells were transfected with gRNA-Cas9 co-expression plasmids for 48 h, and the total genomic DNA was extracted for PCR analysis withLMP1-(up) and LMP2A-specific (down) primers. e and f: Real-time quantitative PCR analysis (e) and western blotting (f) of LMP1 and LMP2A in ‘LMP1 up’ CNE-2 cells that were transfected with LMP1- or LMP2A-specific gRNA-Cas9 co-expression plasmids for 48 (for mRNA quantification) or 72 (for western blotting) hours. Experiments were independently performed in triplicate. *p < 0.05; ***p < 0.001; **** p < 0.0001; ns, no significance