Stable inhibition of hepatitis B virus expression and replication by expressed siRNA

RNA interference might be an efficient antiviral therapy for some obstinate illness. Here, we studied the effects of hepatitis B virus (HBV)-specific 21-nt small interfering RNAs (siRNA) on HBV gene expression and replication in 2.2.15 cells. Seven vectors expressing specific hairpin siRNA driven by...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2005-10, Vol.335 (4), p.1051-1059
Main Authors: Ren, Guang-Li, Bai, Xue-Fan, Zhang, Yan, Chen, Hong-Mei, Huang, Chang-Xing, Wang, Ping-Zhong, Li, Guang-Yu, Zhang, Ying, Lian, Jian-Qi
Format: Article
Language:English
Subjects:
Antiviral therapy
Cell Line, Tumor
Gene Expression Regulation, Viral - genetics
Gene Silencing
Genomic Instability - genetics
Hepatitis B virus
Hepatitis B virus - genetics
Hepatitis B virus - growth & development
Hepatoblastoma - virology
Humans
Liver Neoplasms - virology
Polymerase II-promoter
RNA interference
RNA, Small Interfering - genetics
Small interfering RNA
Transfection
Virus Replication - genetics
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Summary:RNA interference might be an efficient antiviral therapy for some obstinate illness. Here, we studied the effects of hepatitis B virus (HBV)-specific 21-nt small interfering RNAs (siRNA) on HBV gene expression and replication in 2.2.15 cells. Seven vectors expressing specific hairpin siRNA driven by the RNA polymerase II-promoter were constructed and transfected into 2.2.15 cells. In the cell strain that can stably express functional siRNA, the HBV surface antigen (HBsAg) and the HBV e antigen (HBeAg) secretion into culture media was inhibited by 86% and 91%, respectively, as shown by an enzyme-linked immunosorbent assay. Immunofluorescence and Western blot indicated similar results. HBV DNA was markedly restrained by 3.28-fold, as assessed by the fluorescent quantitation PCR. Moreover, the HBV mRNA was significantly reduced by 80% based on semiquantitative RT-PCR. In conclusion, the specific siRNA can knock down the HBV gene expression and replication in vitro, and the silence effects have no relationship with interferon response.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2005.07.170