Biotechnology Letters

, Volume 28, Issue 20, pp 1679–1685 | Cite as

A retrovirus-based system to stably silence hepatitis B virus genes by RNA interference

  • Fang Jia
  • Yi-Zheng Zhang
  • Chang-Mei LiuEmail author
Original Paper


RNA interference (RNAi) might be an efficient antiviral therapy for some obstinate illness. Herein, a retrovirus-based RNAi system was developed to drive expression and delivery of Hepatitis B virus (HBV)-specific short hairpin RNA (shRNA) in HepG2 cells. The levels of HBsAg and HBeAg and that of HBV mRNA were dramatically decreased by this RNAi system in HepG2 cells transfected with Topo-HBV plasmid. Retrovirus-based RNAi thus may be useful for therapy in HBV and other viral infections and provide new clues for prophylactic vaccine development.


Hepatitis B virus HepG2 cells Retrovirus vector RNA interference 


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We thank Dr. David R. Engelke of the University of Michigan, USA for providing the plasmid pAVU6 + 27 and Dr. DePei Liu of Peking Union Medical College, China for providing the plasmid pXSN and pXRN. We also thank Fuliang Chu for creating the figures. This work was supported by grants from National Projects 973 (Grants No. 2005CB522901 and 2001CB510001). Fang Jia is as co-training Ph.D student by the School of Life Sciences, Sichuan University (SLS) and Molecular Virology Research Center, Institute of Microbiology, CAS (MVRC). Her research work was directed and carried out in the MVRC. Therefore SLS and MVRC share the first institution.


  1. Barton GM, Medzhitov R (2002) Retroviral delivery of small interfering RNA into primary cells. Proc Natl Acad Sci USA 99:14943–14945PubMedCrossRefGoogle Scholar
  2. Bridge AJ, Pebernard S, Ducraux A, Nicoulaz AL, Iggo R (2003) Induction of an interferon response by RNAi vectors in mammalian cells. Nat Genet 34:263–264PubMedCrossRefGoogle Scholar
  3. Brummelkamp TR, Bernards R, Agami R (2002) A system for stable expression of short interfering RNAs in mammalian cells. Science 296:550–553PubMedCrossRefGoogle Scholar
  4. Capodici J, Kariko K, Weissman D (2002) Inhibition of HIV-1 infection by small interfering RNA-mediated RNA interference. J Immunol 169:5196–5201PubMedGoogle Scholar
  5. Dong W, Liu D, Li J, Li B, Guo Z, Liang CC (2000) Comparison of two kinds of methods to determine the titer of recombinant retrovirus containing beta-globin gene based on G418 selection. Mol Biotechnol 15:77–81PubMedCrossRefGoogle Scholar
  6. Ge Q, McManus MT, Nguyen T, Shen CH, Sharp PA, Eisen HN, Chen J (2003) RNA interference of influenza virus production by directly targeting mRNA for degradation and indirectly inhibiting all viral RNA transcription. Proc Natl Acad Sci USA 100:2718–2723PubMedCrossRefGoogle Scholar
  7. Giladi H, Ketzinel-Gilad M, Rivkin L, Felig Y, Nussbaum O, Galun E (2003) Small interfering RNA inhibits hepatitis B virus replication in mice. Mol Ther 8:769–776PubMedCrossRefGoogle Scholar
  8. Gitlin L, Karelsky S, Andino R (2002) Short interfering RNA confers intracellular antiviral immunity in human cells. Nature 418:430–434PubMedCrossRefGoogle Scholar
  9. Hammond SM, Bernstein E, Beach D, Hannon GJ (2000) An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells. Nature 404:293–296PubMedCrossRefGoogle Scholar
  10. Hannon GJ (2002) RNA interference. Nature 418:244–251PubMedCrossRefGoogle Scholar
  11. Konishi M, Wu CH, Wu GY (2003) Inhibition of HBV replication by siRNA in a stable HBV-producing cell line. Hepatology 38:842–850PubMedGoogle Scholar
  12. McCaffrey AP, Nakai H, Pandey K, Huang Z, Salazar FH, Xu H, Wieland SF, Marion PL, Kay MA (2003) Inhibition of hepatitis B virus in mice by RNA interference. Nat Biotechnol 21:639–644PubMedCrossRefGoogle Scholar
  13. Randall G, Grakoui A, Rice CM (2003) Clearance of replicating hepatitis C virus replicon RNAs in cell culture by small interfering RNAs. Proc Natl Acad Sci USA 100:235–240PubMedCrossRefGoogle Scholar
  14. Ren GL, Bai XF, Zhang Y, Chen HM, Huang CX, Wang PZ, Li GY, Zhang Y, Lian JQ (2005) Stable inhibition of hepatitis B virus expression and replication by expressed siRNA. Biochem Biophys Res Commun 335:1051–1059PubMedCrossRefGoogle Scholar
  15. Rubinson DA, Dillon CP, Kwiatkowski AV, Sievers C, Yang L, Kopinja J, Rooney DL, Ihrig MM, McManus MT, Gertler FB, et al. (2003) A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference. Nat Genet 33:401–406PubMedCrossRefGoogle Scholar
  16. Shlomai A, Shaul Y (2003) Inhibition of hepatitis B virus expression and replication by RNA interference. Hepatology 37:764–770PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  1. 1.School of Life SciencesSichuan University Chengdu China
  2. 2.Molecular Virology Research Center, Institute of MicrobiologyChinese Academy of Sciences Beijing China

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