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| Inhibition of Hepatitis B Virus by Small Interfering RNA Expressed from Adenovirus-associated Virus Vectors |
| XIN Juan-juan1,MENG Qing-ling2,GAO Xiang-dong1,LIU Jin-yi3 |
1.School of life Science & Technology, China Pharmaceutical University, Nanjing 210009,China
2.Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
3.Beijing Tri-Prime Genetic Engineering Co. Ltd, Beijing102600, China |
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Abstract Objective: To construct the recombinant Adenovirus-associated virus(AAV) vector for siRNA targeting HBV mRNA and to evaluate the inhibitive effect on Hep G 2.2.15 cell. Method: The packing cell line(human embryonic kidney 293T cell) was co-transfected with AAV backbone plasmid pAAV-MCS which has been cloned into the selected siRNA , along with the pAAV-RC and pHelper in AAV Helper-Free System. The recombinant Adenovirus-associated virus was packaged and amplified, followed by infection of the Hep G 2.2.15 cells. The expression level of HBV gene and the replication of HBV were analyzed by ELISA and fluorescence quantitative RCR. Result: The recombinant Adenovirus-associated virus vectors containing siRNA targeting HBV mRNA was successfully constructed. ELISA results demonstrated that the siRNA can effectively inhibit the secretion of HBsAg and HBeAg and the fluorescence quantitative RCR results also showed that the copies of HBV DNA and RNA were significantly reduced. Conclusion: The recombinant Adenovirus-associated virus vector can effectively inhibit the expression and replication of HBV in vitro.
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Received: 26 February 2010
Published: 29 April 2010
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