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PMID: 8628671 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Regulation of Cre recombinase activity by the synthetic steroid RU 486.

Nucleic acids research ·Vol. 24 ·No. 8 ·1996-04-15 ·Pages 1404-11

Kellendonk C, Tronche F, Monaghan AP, Angrand PO, Stewart F, Schütz G

Abstract

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.

MeSH Terms
Animals Base Sequence Cell Line Chlorocebus aethiops DNA Nucleotidyltransferases/drug effects,genetics,metabolism Escherichia coli Gene Expression Humans Integrases Mice Mifepristone/pharmacology Molecular Sequence Data Oligodeoxyribonucleotides Receptors, Progesterone/genetics,metabolism Recombinant Fusion Proteins/genetics Recombination, Genetic Stem Cells Viral Proteins
Chemicals
Oligodeoxyribonucleotides Receptors, Progesterone Recombinant Fusion Proteins Viral Proteins Mifepristone Cre recombinase DNA Nucleotidyltransferases Integrases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Kellendonk C
Molecular Biology of the Cell 1, German Cancer Research Center, Heidelberg.
Tronche F
Monaghan A P
Angrand P O
Stewart F
Schütz G
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/24.8.1404
Published
1996-04-15
Pages
1404-11
Language
English
Region
England
NLM ID
0411011
PMCID
PMC145830
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/24.8.1404 PMC Full Text View journal details
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