Dual gRNAs guided CRISPR/Cas9 system inhibits hepatitis B virus replication

Figure 2 Fifteen hepatitis B virus-specific gRNAs could efficiently suppress the production of hepatitis B virus surface antigen or hepatitis B virus e antigen. A: The plasmid pBB4.5-HBV1.2 (0.5 μg) was co-transfected with each individual gRNA/Cas9 dual expression vector (1.5 μg) to HuH-7 cells. HBsAg level in culture supernatant was measured at 72 h post transfection using a time-resolved fluoroimmunoassay analysis; B: HBeAg level in culture supernatant was measured using a time-resolved fluoroimmunoassay analysis as above; C: The cytotoxicity of each HBV-specific gRNA was examined using an MTT assay. Data are shown as mean ± SE of 5 independent experiments. All P-values are from Student’s t-test. HBV: Hepatitis B virus; HBsAg: Hepatitis B virus surface antigen; HBeAg: Hepatitis B virus e antigen; MTT: Methyl thiazolyl tetrazolium. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001.