Temporal control of gene expression in transgenic mice by a tetracycline-responsive promoter.
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 beta-galactosidase reporter gene under the control of PhCMV*-1 or 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 beta-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.
Furth
P A
PA
Department of Molecular Cell Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
St Onge
L
L
Böger
H
H
Gruss
P
P
Gossen
M
M
Kistner
A
A
Bujard
H
H
Hennighausen
L
L
D000818:Animals; D001483:Base Sequence; D003587:Cytomegalovirus / Q000235:genetics; D017931:DNA Primers; D004251:DNA Transposable Elements; D004622:Embryo, Mammalian / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005865:Gestational Age; D006801:Humans; D008156:Luciferases / Q000032:analysis / Q000096:biosynthesis; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009132:Muscles / Q000378:metabolism; D009876:Operon; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000187:drug effects; D018139:Simplexvirus / Q000235:genetics; D013752:Tetracycline / Q000494:pharmacology*; D013997:Time Factors; D014059:Tongue / Q000378:metabolism; D015534:Trans-Activators / Q000096:biosynthesis*; D001616:beta-Galactosidase / Q000032:analysis / Q000096:biosynthesis
D017931:DNA Primers; D004251:DNA Transposable Elements; D015534:Trans-Activators; D008156:Luciferases; D001616:beta-Galactosidase; D013752:Tetracycline