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Am J Physiol Renal Physiol (August 19, 2003). doi:10.1152/ajprenal.00301.2002
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Submitted on August 20, 2002
Accepted on July 27, 2003

Conditional gene expression in renal collecting duct epithelial cells : Use of the inducible Cre-Lox system

Antoine Ouvrard-Pascaud1, Stefania Puttini1, Yannis Sainte-Marie1, Rafika Athman2, Virginie Fontaine1, Francoise Cluzeaud1, Nicolette Farman1, Marie-Edith Rafestin-Oblin1, Marcel Blot-Chabaud1, and Frederic Jaisser1*

1 INSERM U478, Institut National de la Sante et de la Recherche Medicale, PARIS, France
2 UMR 144, CNRS, Institut Curie, PARIS, France

* To whom correspondence should be addressed. E-mail: frederic.jaisser{at}college-de-france.fr.

The renal collecting duct (CD) plays a key role in the control of ion and fluid homeostasis. Genes encoding for ion transporters, hormone receptors or regulatory proteins specifically expressed in CD are mutated in several genetic diseases with altered blood pressure. Suitable cellular models expressing genes in a conditional way should represent attractive systems for structure-function relationship analyses and the generation of appropriate physiopathological models of related diseases. However, the generation of such systems remains laborious and quite inefficient. We adapted and improved a conditional Cre-Lox inducible system in the highly differentiated aldosterone-sensitive rat collecting duct RCCD2 cell line. The inducible MerCreMer recombinase allowed tight control and high levels of transgene expression, while flanking a selection marker with two LoxP sites strongly improved the selection procedure. We have used this system to conditionally express an eGFP-tagged human mineralocorticoid receptor (eGFP-hMR). This will allow in the future structure-function analyses as well as MR trafficking studies in these epithelial cells that retain the features of the native collecting duct. Of note, improvements of the conditional Cre-Lox expression system have potentially wide applications in other epithelial or non-epithelial cell lines.




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