RNA Interference Mediated The Inhibition Of HBV Replication And Expression And Its Mechanisms Research

Objective: To observe the inhibition of HBV replication and expression by RNA interference, and try to investigate the function of Dicer in RNA interference.Methods: The eukaryotic expression plasmid pHBV, which contained 1.3-fold-overlength genome of HBV, were constructed and transfected into HepG2 cells so that made a model of HBV infection, then evaluated by ELISA, immunofluorescence staining, Southern and Northern blot; Constructed ten kinds of shRNAs expression vectors, which aimed at four open read frames of HBV, and cotransfected them with pHBV into HepG2 cells, then observed by ELISA, immunofluorescence staining, Southern, Northern and Western blot,RT-PCR;Constructed two kinds of shRNAs expression vectors named pShRNA-D1 and pShRNA-D2,which aimed at the RNAaseâ…¢of Dicer gene, then transfected them into HepG2 2.2.15 cells, Colon Cancer TC cells and HepG2 A9 cells, which intergrated GFP(Green Fluorescent Protein) gene, the level of Dicer mRNA was investigated by RT-PCR. After knocked down the Dicer gene of HepG2 A9...