Targeted Screening Of SiRNA Directed HBV Polymerase Gene For Effectively Inhibiting HBV Expression

AIM: To screen potential mRNA locations against which targeting siRNAs can effectively inhibit HBV expression.METHODS: Five recombinant plasmids containing four targeting-specific siRNA fragments and a control were prepared and transfected into 2.2.15 respectively. The expression levels of HBs mRNA and HBc mRNA, HBx mRNA, mRNAs were detected by RT-PCR, respectively. The concentrations of the hepatitis B virus antigens, including HBsAg and HBeAg harvested from the culture supernatant of transfected 2.2.15 cells, were measured by ELISA. X protein was tested by Western Blot.RESULTS: Four siRNAs against distinct mRNA locations of HBV polymerse gene, shown different inhibitory effects on their targeted mRNA. The plasmid-derived psiRNA1 and psiRNA2 could effectively inhibit transcription and translation of HBs gene, whereas the inhibitory efficiency of psiRNA3, psiRNA4 for HBe gene is much higher than that of psiRNA1and psiRNA2. Comparing to rest of psiRNAs in this study, psiRNA4 is the most effective to suppress the transcription and translation of HBx.CONCLUSION: siRNA can be considered as a powerful therapeutic agent for reducing HBV replication and translation. The siRNAs for HBV polymerase is effective largely depending on the location of targeted sites. To enhance inhibitory efficiency, hunting for high effective target in polymerase gene is necessary and feasible.