[Inhibition of HBV replication and antigen expression by RNA interference against different targets].
Bingqiang Zhang, Ying Huang, Ni Tang, Ying Wu, Jun Zhang, Weixian Chen, Tong‐Chuan He, Ailong Huang
Abstract
Bingqiang Zhang, Ying Huang, Ni Tang, Ying Wu, Jun Zhang, Weixian Chen, Tong‐Chuan He, Ailong Huang
Abstract
OBJECTIVES: To observe the inhibition of HBV replication and antigen expression by RNA interference aimed at different parts of the HBV genome. METHODS: Following the rules of shRNA expression vector design and construction, we constructed seven kinds of sequence specific vectors and two kinds of mutant shRNA expression ones. We then cotransfected those shRNA and HBV expression vectors into HepG2 cells using lipofectamine2000. The level of HBV replication was investigated using Southern blot and the antigen expression using ELISA. RESULTS: The replication of HBV DNA was inhibited by many shRNAs, especially the ones against P1, S2, C2, S1 and X. The inhibition rate against P1 was as high as 95%. Results obtained with ELISA showed that the shRNAs targeting C2, C1 and S2 had high rates of inhibition to HBsAg. CONCLUSION: The replication and antigen expression of HBV could be inhibited by shRNAs aimed at four different open read frames, and higher inhibition rates of HBV replication and surface antigen expression could be obtained by P1 and C2, respectively.
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OBJECTIVES: To observe the inhibition of HBV replication and antigen expression by RNA interference aimed at different parts of the HBV genome. METHODS: Following the rules of shRNA expression vector design and construction, we constructed seven kinds of sequence specific vectors and two kinds of mutant shRNA expression ones. We then cotransfected those shRNA and HBV expression vectors into HepG2 cells using lipofectamine2000. The level of HBV replication was investigated using Southern blot and the antigen expression using ELISA. RESULTS: The replication of HBV DNA was inhibited by many shRNAs, especially the ones against P1, S2, C2, S1 and X. The inhibition rate against P1 was as high as 95%. Results obtained with ELISA showed that the shRNAs targeting C2, C1 and S2 had high rates of inhibition to HBsAg. CONCLUSION: The replication and antigen expression of HBV could be inhibited by shRNAs aimed at four different open read frames, and higher inhibition rates of HBV replication and surface antigen expression could be obtained by P1 and C2, respectively.
Key concepts: Small hairpin RNA, HBsAg, Lipofectamine, RNA interference, Virology, Molecular biology, Biology, Hepatitis B virus