Inhibition of NF-kappaB mediated inflammation by siRNA expressed by recombinant adeno-associated virus.
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.
Pinkenburg
Olaf
O
Department of Internal Medicine, Division of Pulmonary Diseases, Hospital of the University of Marburg, Philipps-Universtät Marburg, Baldingerstrasse 1, D-35043 Marburg, Germany.
Platz
Juliane
J
Beisswenger
Christoph
C
Vogelmeier
Claus
C
Bals
Robert
R
D002460:Cell Line; D000229:Dependovirus / Q000235:genetics*; D020868:Gene Silencing; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism*; D016209:Interleukin-8 / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics* / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics*; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology
D018836:Inflammation Mediators; D016209:Interleukin-8; D016328:NF-kappa B; D034741:RNA, Small Interfering; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; D008156:Luciferases