Control of cytokine gene expression using small RNA interference: blockade of interleukin-10 and interferon-gamma gene expression in pig cells.
The ability of small RNA interference (RNAi) to reduce specific gene expression was tested using interleukin-10 (IL-10) and interferon-gamma (IFN-gamma) production by cultured swine blood mononuclear cells stimulated by Escherichia coli lipopolysaccharide or concanavalin A. Antisense (AS) phosphorothioate oligodeoxynucleotides (ODNs) corresponding to a sequence in the region of the AUG initiation codon of swine IL-10 or IFN-gamma mRNA inhibited production of IL-10 (>or=93.5%) and IFN-gamma (>or=99%) mRNAs. Interleukin-10 and IFN-gamma protein production was inhibited more than 95% by the AS ODNs. Scrambled and sense ODNs RNAi used as negative controls did not alter mRNA expression for either cytokine but slightly reduced IL-10 protein production. Cytokine-specific and control RNAi did not inhibit beta(2)-microglobulin mRNA expression in mitogen-stimulated blood mononuclear cells. Thus AS ODNs RNAi specifically inhibit expression of pig IL-10 and IFN-gamma mRNAs by cultured, mitogen-stimulated blood mononuclear cells and may be an attractive alternative method for studying cytokine function.
Sidahmed
A M E
AM
Department of Pathobiology, University of Guelph, Guelph, Ontario, Canada N1G 2W1.
Wilkie
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BN
D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005786:Gene Expression Regulation / Q000235:genetics / Q000276:immunology*; D007371:Interferon-gamma / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D016753:Interleukin-10 / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000502:physiology; D009841:Oligonucleotides / Q000276:immunology; D012313:RNA / Q000737:chemistry / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D013047:Specific Pathogen-Free Organisms; D013552:Swine / Q000235:genetics / Q000276:immunology*; D001613:beta 2-Microglobulin / Q000235:genetics / Q000276:immunology
D009841:Oligonucleotides; D034741:RNA, Small Interfering; D001613:beta 2-Microglobulin; D016753:Interleukin-10; D012313:RNA; D007371:Interferon-gamma