tailieunhanh - Báo cáo khoa học: Effect of ecdysone receptor gene switch ligands on endogenous gene expression in 293 cells

Regulated gene expression may substantially enhance gene therapy. Corre-lated with structural differences between insect ecdysteroids and mamma-lian steroids, the ecdysteroids appear to have a benign pharmacology without adversely interfering with mammalian signaling systems. Conse-quently, the ecdysone receptor-based gene switches are attractive for appli-cation in medicine. | ỊFEBS Journal Effect of ecdysone receptor gene switch ligands on endogenous gene expression in 293 cells Siva K. Panguluri1 Bing Li2 Robert E. Hermann2 and Subba R. Palli1 1 Department of Entomology College of Agriculture University of Kentucky Lexington KY USA 2 Intrexon Corporation Norristown PA USA Keywords diacylhydrazine ecdysone gene therapy microarray RSL-1 Correspondence S. R. Palli Department of Entomology College of Agriculture University of Kentucky Lexington KY 40546 USA Fax 1 859 323 1120 Tel 1 859 257 4962 E-mail rpalli@ Present address MicroBiotiX Inc. Worcester MA USA Received 24 June 2007 revised 13 August 2007 accepted 4 September 2007 doi Regulated gene expression may substantially enhance gene therapy. Correlated with structural differences between insect ecdysteroids and mammalian steroids the ecdysteroids appear to have a benign pharmacology without adversely interfering with mammalian signaling systems. Consequently the ecdysone receptor-based gene switches are attractive for application in medicine. In the present study the effect of inducers of ecdysone receptor switches on the expression of endogenous genes in HEK 293 cells was determined. Four ligand chemotypes represented by a tetrahydroquinoline RG-120499 one amidoketone RG-121150 two ecdysteroids 20-hydroxyecdysone 20E and ponasterone A Pon A and four diacylhydrazines rG-102240 RG-1o2277 RG-102398 and RG-100864 were tested in HEK 293 cells. The cells were exposed to ligands at concentrations of 1 pM RG-120499 or 10 pM all others for 72 h and the total RNA was isolated and analyzed using microarrays. Microarray data showed that the tetrahydroquinoline ligand RG-120499 caused cell death at concentrations 10 pM. At 1 pM this ligand caused changes in the expression of genes such as TNF MAF Rab and Reprimo. At 10 pM the amidoketone RG-121150 induced changes in the expression of genes such as v-jun FBJ and EGR but was otherwise noninterfering. Of the .