Generation of conditional mutants in higher eukaryotes by switching between the expression of two genes
- Udo Baron*,
- Dirk Schnappinger†,
- Vera Helbl†,
- Manfred Gossen‡,
- Wolfgang Hillen†, and
- Hermann Bujard*,§
- *Zentrum für Molekulare Biologie, Universität Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany; †Lehrstuhl für Mikrobiologie, Universität Erlangen, Staudtstrasse 5, D-91058 Erlangen, Germany; and ‡University of California, MCB Barker/Koshland ASU, 401 Barker Hall, Berkeley, CA 94720-3202
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Edited by Eric R. Kandel, Columbia University College of Physicians and Surgeons, New York, NY, and approved November 5, 1998 (received for review September 28, 1998)
Abstract
A regulatory system for the in-depth study of gene functions in higher eukaryotic cells has been developed. It is based on the tetracycline-controlled transactivators and reverse tTA, which were remodeled to discriminate efficiently between two different promoters. The system permits one to control reversibly the activity of two genes, or two alleles of a gene, in a mutually exclusive way, and also allows one to abrogate the activities of both. This dual regulatory circuit, which can be operated by a single effector substance such as doxycycline, overcomes limitations of conventional genetic approaches. The conditional mutants that can now be generated will be useful for the study of gene function in vitro and in vivo. In addition, the system may be of value for a variety of practical applications, including gene therapy.
Footnotes
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↵ § To whom reprint requests should be addressed. e-mail: bujardh{at}sun0.urz.uni-heidelberg.de.
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This paper was submitted directly (Track II) to the Proceedings Office.
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A Commentary on this article begins on page 797.
- ABBREVIATIONS:
- tTA,
- tetracycline-controlled transactivator;
- rtTA,
- reverse tTA;
- Tc,
- tetracycline;
- Dox,
- doxycycline;
- ATc,
- anhydrotetracycline
- Copyright © 1999, The National Academy of Sciences
