The Specific Inhibitory Effect of shRNA Targeting X Gene on Hepatitis B Virus Gene Expression and Replication

Objective To investigate the inhibitory effect on Hepatitis B virus(HBV) gene expression and replication by small hairpin RNAs(shRNA) targeting X gene.Methods Four eukaryotic expression plasmids(pshHBx1-4) were constructed to express shRNAs targeting HBV X gene with molecular cloning methods.HepG2 cells were cotransfected with the shRNA expression plasmids and a replication-competent HBV construct.The mRNA of HBV and replicative intermediates were determined with Northern blot and Southern blot,the expression of HBcAg was analyzed with immunofluorescence staining.Results Cotransfection of pshHBx1-4 was significantly inhibited the expression of HBV mRNA,decreased the expression levels of HBsAg and HBeAg in cell culture supernatant and intracellular HBcAg,effectively inhibited the synthesis of HBV replicative intermediates.Conclusion X-gene specific shRNAs can effectively inhibit HBV gene expression and replication and may develop a novel antiviral agent for HBV infection treatment.