[Inhibition of HBV core antigen gene expression in human embryonic kidney cell line AD293 by plasmid-based RNAi]

Zhejiang Da Xue Xue Bao Yi Xue Ban. 2005 Mar;34(2):104-9. doi: 10.3785/j.issn.1008-9292.2005.02.003.
[Article in Chinese]

Abstract

Objective: To inhibit HBV core antigen gene expression with plasmid-based RNAi.

Methods: The shRNA expression vector targeting HBV core antigen gene was designed and constructed. Human embryonic kidney cell line AD293 was co-transfected with HBcAg-EGFP fusion protein expression vector and shRNA expression vector transiently, and the cells without shRNA-transfection and with non-specific shRNA transfection were used as controls. Inhibitory effect of RNAi was detected by fluorescence-activated cell sorting (FACS) and real-time fluorescence quantificational RT-PCR.

Results: HBV core antigen gene expression in AD293 was inhibited by shRNA, with the maximal inhibition rate of 76 % measured by FACS and of 63.1 % by real-time PCR.

Conclusion: Effective inhibition of HBV core antigen gene expression by plasmid-based RNAi provides an alternative for anti-HBV study in vitro, which has potential clinical application.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Embryo, Mammalian
  • Gene Expression Regulation, Viral*
  • Green Fluorescent Proteins / biosynthesis
  • Green Fluorescent Proteins / genetics
  • Hepatitis B Core Antigens / biosynthesis*
  • Hepatitis B Core Antigens / genetics
  • Humans
  • Kidney / cytology
  • Kidney / virology*
  • Plasmids
  • Polymerase Chain Reaction
  • RNA Interference / physiology*
  • RNA, Messenger / genetics
  • Transfection

Substances

  • Hepatitis B Core Antigens
  • RNA, Messenger
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins