Regulation of Cre Recombinase Activity by the Synthetic Steroid RU 486

To create a strategy for inducible gene targeting we developed a Cre-lox recombination system which responds to the synthetic steroid RU 486. Several fusions between Cre recombinase and the hormone binding domain (HBD) of a mutated human progesterone receptor, which binds RU 486 but not progesterone...

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Bibliographic Details
Published in:Nucleic acids research 1996-04, Vol.24 (8), p.1404-1411
Main Authors: Kellendonk, Christoph, Tronche, François, Monaghan, A.-Paula, Angrand, Pierre-Olivier, Stewart, Francis, Schütz, Günther
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Cell Line
Cercopithecus aethiops
DNA Nucleotidyltransferases - drug effects
DNA Nucleotidyltransferases - genetics
DNA Nucleotidyltransferases - metabolism
Escherichia coli
Gene Expression
Humans
Integrases
Mice
Mifepristone - pharmacology
Molecular Sequence Data
Oligodeoxyribonucleotides
Receptors, Progesterone - genetics
Receptors, Progesterone - metabolism
Recombinant Fusion Proteins - genetics
Recombination, Genetic
Stem Cells
Viral Proteins
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Summary:To create a strategy for inducible gene targeting we developed a Cre-lox recombination system which responds to the synthetic steroid RU 486. Several fusions between Cre recombinase and the hormone binding domain (HBD) of a mutated human progesterone receptor, which binds RU 486 but not progesterone, were constructed. When tested in transient expression assays recombination activities of all fusion proteins were responsive to RU 486, but not to the endogenous steroid progesterone. However, the observed induction of recombination activity by the synthetic steroid varied between the different fusion proteins. The fusion with the highest activity in the presence of RU 486 combined with low background activity in the absence of the steroid was tested after stable expression in fibroblast and embryonal stem (ES) cells. We could demonstrate that its recombination activity was highly dependent on RU 486. Since the RU 486 doses required to activate recombination were considerably lower than doses displaying anti-progesterone effects in mice, this system could be used as a valuable tool for inducible gene targeting.
ISSN:0305-1048
1362-4962
1362-4962
DOI:10.1093/nar/24.8.1404