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

Journal of Virological Methods
Olaf PinkenburgRobert Bals

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.

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Citations

Jun 14, 2013·Evidence-based Complementary and Alternative Medicine : ECAM·Wang-Jian ZhaMao Huang
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Oct 5, 2010·The Journal of Immunology : Official Journal of the American Association of Immunologists·Wei LiYing Zhu

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