Published online Jul 1, 2004. doi: 10.3748/wjg.v10.i13.1898
Revised: January 8, 2004
Accepted: January 15, 2004
Published online: July 1, 2004
AIM: To study the effect of siRNA expressed from DNA vector on HBV replication.
METHODS: Human U6 promoter was amplified from genomic DNA and cloned into plasmid pUC18 to construct a mammalian siRNA expression vector pUC18U6. Then oligonucleotides coding for a short hairpin RNA against HBV were cloned into pUC18U6 to form pUC18U6HBVsir which was introduced into 2.2.15 cells by using liposome-mediated transfection. 2.2.15 cells transfected by pUC18U6 and pUC18U6GFPsir which expressed siRNA against green fluorescent protein and mock-transfected 2.2.15 cells were used as controls. Concentration of HBsAg in the supernatant of the transfected cells was measured by using solid-phase radioimmunoassay.
RESULTS: A mammalian siRNA expression vector pUC18U6 was constructed successfully. Compared with controls, pUC18U6HBVsir which expressed siRNA against HBV decreased concentration of HBsAg significantly by 44% (P < 0.05).
CONCLUSION: HBV replication in 2.2.15 cells is inhibited by siRNA expressed from the DNA vector.