Keratinocyte gene therapy: inducible promoters and in vivo control of transgene expression.
Modulation of transgene expression by exogenous agents is an optimal goal in gene therapy. Successful keratinocyte gene therapy requires a promoter-enhancer cassette to regulate expression of the therapeutic gene in vivo. In this study, we first transferred plasmids, constructed by introducing inducible promoters fused to the beta-galactosidase gene (LAC Z), into keratinocytes in vitro. Metallothionein (MT) and 1,24-vitamin D(3)(OH)(2) dehydroxylase (VDH) promoters responded to the inducing agents, Cadmium and 1,25-vitamin D(3)(OH)(2) (VitD(3)), respectively. The plasmids were then introduced in vivo using a naked DNA method and the inducible promoters were evaluated by measuring beta-gal activity in rat keratinocytes. Zinc induced the transferred MT promoter activity by approximately 2-fold or 10-fold when administered systemically and topically, respectively. In addition, VitD(3) induced the transferred VDH promoter activity approximately 10-fold when administered topically. These data are useful for developing inducible promoters for keratinocyte gene therapy.
Itai
K
K
Department of Dermatology, Hirosaki University School of Medicine, Japan. itai20000@yahoo.co.jp
Sawamura
D
D
Meng
X
X
Hashimoto
I
I
D000818:Animals; D002104:Cadmium / Q000494:pharmacology; D002117:Calcitriol / Q000494:pharmacology; D002460:Cell Line; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism*; D008668:Metallothionein / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D051381:Rats; D011919:Rats, Inbred Strains; D011923:Rats, Nude; D019076:Transgenes; D015032:Zinc / Q000494:pharmacology; D001616:beta-Galactosidase / Q000235:genetics
D002104:Cadmium; D008668:Metallothionein; D001616:beta-Galactosidase; D002117:Calcitriol; D015032:Zinc