Temporal Control of Gene Expression in Transgenic Mice by a Tetracycline- Responsive Promoter

• Published in: Proceedings of the National Academy of Sciences - PNAS 1994-09, Vol.91 (20), p.9302-9306
• Main Authors: Furth, Priscilla A., St. Onge, Luc, Böger, Hinrich, Gruss, Peter, Gossen, Manfred, Kistner, Andreas, Bujard, Hermann, Hennighausen, Lothar
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; beta-Galactosidase - analysis; beta-Galactosidase - biosynthesis; Biochemistry; Cytomegalovirus - genetics; DNA Primers; DNA Transposable Elements; Embryo, Mammalian - metabolism; Embryos; Enhancer Elements, Genetic; Escherichia coli - genetics; Gene Expression; Gene induction; Genes; Gestational Age; herpes simplex virus; Humans; Luciferases - analysis; Luciferases - biosynthesis; Mice; Mice, Transgenic; Molecular Sequence Data; Muscles - metabolism; Operon; Organ Specificity; Polymerase Chain Reaction; Promoter Regions, Genetic - drug effects; Reporter genes; Rodents; Simplexvirus - genetics; Tetracycline - pharmacology; Thigh; Time Factors; Tongue - metabolism; Trans-Activators - biosynthesis; Transactivators; Transgenes; Transgenic animals
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.91.20.9302

Promoters whose temporal activity can be directly manipulated in transgenic animals provide a tool for the study of gene functions in vivo. We have evaluated a tetracycline-responsive binary system for its ability to temporally control gene expression in transgenic mice. In this system, a tetracycline-controlled trans-activator protein (tTA), composed of the repressor of the tetracycline-resistance operon (tet from Escherichia coli transposon Tn10) and the activating domain of viral protein VP16 of herpes simplex virus, induces transcription from a minimal promoter (PhCMV*-1; see below) fused to seven tet operator sequences in the absence of tetracycline but not in its presence. Transgenic mice were generated that carried either a luciferase or a β-galactosidase reporter gene under the control of PhCMV*-1or a transgene containing the tTA coding sequence under the control of the human cytomegalovirus immediate early gene 1 (hCMV IE1) promoter/enhancer. Whereas little luciferase or β-galactosidase activity was observed in tissues of mice carrying only the reporter genes, the presence of tTA in double-transgenic mice induced expression of the reporter genes up to several thousand-fold. This induction was abrogated to basal levels upon administration of tetracycline. These findings can be used, for example, to design dominant gain-of-function experiments in which temporal control of transgene expression is required.