title: Tetracycline-regulated transactivators driven by the involucrin promoter to achieve epidermal conditional gene expression.
abstract: To achieve conditional gene expression in the differentiated layers of the epidermis, we generated transgenic mice with the tetracycline-regulated transactivator proteins, tTA (tetracycline transactivator) and rtTA (reverse tetracycline transactivator), expressed from the human involucrin promoter. Interaction with tetracycline turns off or turns on the tTA and rtTA molecules, respectively, allowing for regulation of downstream target genes during development and postnatally. These transactivator lines were crossed with reporter mice driving LacZ expression from a tetracycline response element to analyze the specificity and levels of target gene expression. Quantitative beta-galactosidase experiments demonstrate a 30-fold induction, specific to epithelial tissues. Immunohistochemistry results illustrate that the beta-galactosidase staining follows that of endogenous involucrin expression. Induction initiates at embryonic day 14.5 with expression over the entire epidermal surface by E16.5. Together with other driver lines, expressing tetracycline transactivators in the mitotically active layers of the epidermis, these mice will allow investigators to specifically modulate expression of target genes to specific stages of epidermal differentiation.
authors.affiliation: National Human Genome Research Institute, NIH, Bethesda, Maryland 20892, USA.
authors.forename: Jean
authors.initials: J
authors.lastname: Jaubert
authors.forename: Satyakam
authors.initials: S
authors.lastname: Patel
authors.forename: Jun
authors.initials: J
authors.lastname: Cheng
authors.forename: Julia A
authors.initials: JA
authors.lastname: Segre
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004318:Doxycycline / Q000494:pharmacology*; D004817:Epidermis / Q000502:physiology*; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006801:Humans; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D011498:Protein Precursors / Q000235:genetics*; D015533:Transcriptional Activation / Q000187:drug effects / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D011498:Protein Precursors; C032253:involucrin; D001616:beta-Galactosidase; D004318:Doxycycline