Inhibition of hepatitis C virus protein expression by RNA interference

Adrish Sen, Robert Steele, Asish K. Ghosh, Arnab Basu, Ranjit Ray, Ratna B. Ray

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

Hepatitis C virus (HCV) is a serious human pathogen and an estimated 170 million people are infected worldwide. Current therapeutic regimens have shown limited efficacy against selected genotypes of the virus. The phenomenon of RNA interference can be used to selectively block homologous genes post-transcriptionally, and has revolutionized approaches to study gene function. In this report, we have demonstrated that small interfering RNAs (siRNAs) targeted against NS5A of HCV genotype 1a specifically inhibit NS5A RNA and protein expression in a human hepatoma (HepG2) cell line. Expression of endogenous α-actin and the ds-RNA activated serine/threonine kinase-PKR were unaltered, demonstrating that the inhibitory effect observed from siRNA was specific to the HCV NS5A protein. We next examined whether siRNA directed against NS5A could inhibit core protein expression, the first gene product synthesized in virus infected cells due to its localization at the 5′ end of the HCV polyprotein. For this purpose, a full-length cDNA clone from HCV (H77, genotype 1a) was used, and results indicated that the introduction of NS5A targeted siRNA resulted in an inhibition of NS5A and core protein expression. Moreover, we observed that this siRNA effectively inhibited NS5A mediated activation of the IL-8 promoter. Taken together, our results demonstrated that siRNA was effective in inhibiting HCV protein expression, and may have therapeutic potential to limit HCV replication in chronically infected patients.

Original languageEnglish (US)
Pages (from-to)27-35
Number of pages9
JournalVirus Research
Volume96
Issue number1-2
DOIs
StatePublished - Oct 2003
Externally publishedYes

Fingerprint

RNA Interference
Hepacivirus
Small Interfering RNA
Proteins
Genotype
RNA
Viruses
Polyproteins
Protein-Serine-Threonine Kinases
Hep G2 Cells
Virus Replication
Interleukin-8
Genes
Actins
Hepatocellular Carcinoma
Complementary DNA
Clone Cells
Gene Expression
Cell Line
Therapeutics

Keywords

  • HCV
  • NS5A protein
  • RNA interference

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Biology
  • Virology

Cite this

Sen, A., Steele, R., Ghosh, A. K., Basu, A., Ray, R., & Ray, R. B. (2003). Inhibition of hepatitis C virus protein expression by RNA interference. Virus Research, 96(1-2), 27-35. https://doi.org/10.1016/S0168-1702(03)00170-9

Inhibition of hepatitis C virus protein expression by RNA interference. / Sen, Adrish; Steele, Robert; Ghosh, Asish K.; Basu, Arnab; Ray, Ranjit; Ray, Ratna B.

In: Virus Research, Vol. 96, No. 1-2, 10.2003, p. 27-35.

Research output: Contribution to journalArticle

Sen, A, Steele, R, Ghosh, AK, Basu, A, Ray, R & Ray, RB 2003, 'Inhibition of hepatitis C virus protein expression by RNA interference', Virus Research, vol. 96, no. 1-2, pp. 27-35. https://doi.org/10.1016/S0168-1702(03)00170-9
Sen, Adrish ; Steele, Robert ; Ghosh, Asish K. ; Basu, Arnab ; Ray, Ranjit ; Ray, Ratna B. / Inhibition of hepatitis C virus protein expression by RNA interference. In: Virus Research. 2003 ; Vol. 96, No. 1-2. pp. 27-35.
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