Development of packageable replication-defective HBV mutants and experimental study on their anti-HBV effects by expressing antisense RNA / 中华传染病杂志

ABSTRACT

Objective To explore the possibility of using HBV as a gene delivery vector, and to test the anti-HBV effects by intracellular expression of antisense RNA. Methods Two parts of HBV genome were reversedly recombined back into overlength HBV genome, which can produce HBV particle, to express antisense RNA complementary to S or S promoter region respectively. HepG 2.2.15 cell lines were transfected with these constructs and the empty vector pMEP4, then positive clones were selected and mixed in respective groups with hygromycin in the culture medium. HBsAg and HBeAg, which exist in the culture medium, were tested by ELISA method and intracellular HBc related HBV DNA was examined by dot blot hybridization. The existence of recombinant HBV virion in the culture medium was examined by PCR. Results The mean inhibitory rates of HBsAg were (2.74?3.83)%、(66.54?4.45)%(P