Interferon-gamma inhibits transgene expression driven by SV40 or CMV promoters but augments expression driven by the mammalian MHC I promoter.
The use of mammalian gene expression vectors has become increasingly important for transgenics and gene therapy as well as basic research. Essential for the success of these vectors in medical research applications is the proper choice of promoter linked to the gene of interest. Many mammalian expression vectors use promoter elements from pathogenic viruses, including simian virus 40 (SV40) and cytomegalovirus (CMV). Lymphokines produced by the immune response to proteins expressed by these vectors could inhibit further transcription initiation by viral promoters. Our objective was to determine the effect of interferon-gamma (IFN-gamma) on transgene expression driven by a viral SV40 or CMV promoter/enhancer and the mammalian promoter/enhancer for the major histocompatibility complex class I (MHC I) gene. We transfected the luciferase gene driven by these three promoters into 14 cell lines of many tissues and several species. Luciferase assays of transfected cells untreated or treated with IFN-gamma indicated that, although the viral promoters could drive luciferase production in all cell lines tested to greater or lesser levels than the MHC I promoter, treatment with IFN-gamma caused inhibition of transgene expression in most of the cell lines and amplification of the MHC I promoter-driven transgene expression in all cell lines. These data indicate that the SV40 and CMV promoter/enhancers may not be a suitable choice for gene delivery especially for immune response applications or in patients where IFN levels may be elevated. The MHC I promoter/enhancer, on the other hand, may be an ideal transgene promoter for applications involving the immune system.
Harms
J S
JS
Department of Animal Health and Biomedical Sciences, University of Wisconsin-Madison 53706, USA.
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D000818:Animals; D002417:Cattle; D002460:Cell Line / Q000187:drug effects / Q000821:virology; D003587:Cytomegalovirus / Q000235:genetics*; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D015870:Gene Expression / Q000187:drug effects*; D005805:Genes, MHC Class I; D005822:Genetic Vectors / Q000737:chemistry / Q000187:drug effects / Q000235:genetics*; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D007700:Kinetics; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008222:Lymphokines / Q000494:pharmacology; D051379:Mice; D011401:Promoter Regions, Genetic; D051381:Rats; D013539:Simian virus 40 / Q000235:genetics*; D014162:Transfection; D015854:Up-Regulation
D008222:Lymphokines; D007371:Interferon-gamma; D008156:Luciferases