Inhibition of NF-kappaB mediated inflammation by siRNA expressed by recombinant adeno-associated virus

J Virol Methods. 2004 Sep 1;120(1):119-22. doi: 10.1016/j.jviromet.2004.04.007.

Abstract

NF-kappaB mediated inflammation is a key process to many diseases. RNA interference (RNAi) is the specific suppression of genes by short double-stranded RNA. It was the aim of the present study to modify NF-kappaBdependent inflammation by small interfering RNA (siRNA) expressed by recombinant adeno-associated virus (rAAV). To study the kinetics of rAAV mediated expression of siRNA, the expression of the luciferase gene was targeted and resulted in a significant decrease of luciferase activity as compared to a control vector in the human 293 cell line. The effect was dose dependent and was detectable 24 h after infection. rAAV coding for siRNA against the p65 subunit of NF-kappaBsignificantly reduced the p65 protein. In a cellular model of TNF-alpha induced inflammation, expression of siRNA against p65 significantly suppressed the secretion of IL-8 from BEAS-2B cells. In conclusion, rAAV vectors coding for siRNA are an useful tool for efficient gene silencing in mammalian cells and can be used to modify NF-kappaB mediated inflammation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Dependovirus / genetics*
  • Gene Silencing*
  • Genes, Reporter
  • Genetic Vectors
  • Humans
  • Inflammation Mediators / metabolism*
  • Interleukin-8 / metabolism
  • Luciferases / genetics
  • Luciferases / metabolism
  • NF-kappa B / genetics*
  • NF-kappa B / physiology*
  • RNA, Small Interfering / genetics*
  • Transcription Factor RelA
  • Tumor Necrosis Factor-alpha / physiology

Substances

  • Inflammation Mediators
  • Interleukin-8
  • NF-kappa B
  • RNA, Small Interfering
  • Transcription Factor RelA
  • Tumor Necrosis Factor-alpha
  • Luciferases