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Generation of conditional mutants in higher eukaryotes by switching between the expression of two genes.

TitleGeneration of conditional mutants in higher eukaryotes by switching between the expression of two genes.
Publication TypeJournal Article
Year of Publication1999
AuthorsBaron U, Schnappinger D, Helbl V, Gossen M, Hillen W, Bujard H
JournalProc Natl Acad Sci U S A
Volume96
Issue3
Pagination1013-8
Date Published1999 Feb 02
ISSN0027-8424
KeywordsBacterial Proteins, DNA-Binding Proteins, Doxorubicin, Gene Expression Regulation, Bacterial, Genes, Switch, HeLa Cells, Humans, Phenotype, Promoter Regions, Genetic, Recombinant Fusion Proteins, Tetracycline, Trans-Activators, Transcriptional Activation, Transfection
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.

DOI10.1073/pnas.96.3.1013
Alternate JournalProc Natl Acad Sci U S A
PubMed ID9927685
PubMed Central IDPMC15342

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