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Nucleic Acids Research 2005 33(5):e51; doi:10.1093/nar/gni051
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Published online 22 March 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
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Conditional and inducible transgene expression in mice through the combinatorial use of Cre-mediated recombination and tetracycline induction

Gusztav Belteki1, Jody Haigh1, Nikolett Kabacs1, Katharina Haigh1, Karen Sison1, Frank Costantini2, Jeff Whitsett3, Susan E. Quaggin1,4 and Andras Nagy1,5,*

1Samuel Lunenfeld Research Institute, Mount Sinai Hospital 600 University Avenue, Toronto, Ontario, Canada M5G 1X5 2Department of Genetics and Development, College of Physicians and Surgeons, Columbia University New York, NY 10032, USA 3Children's Hospital Medical Center, Division of Pulmonary Biology Cincinnati, OH 45229-3039, USA 4St Michael's Hospital Toronto, Ontario, Canada 5Department of Molecular and Medical Genetics, University of Toronto Toronto, Ontario, Canada M5S 1A8

*To whom correspondence should be addressed. Tel: +1 416 586 3246; Fax: +1 416 586 8588; Email: nagy{at}mshri.on.ca

Received January 20, 2005. Revised March 1, 2005. Accepted March 1, 2005.

Here we describe a triple transgenic mouse system, which combines the tissue specificity of any Cre-transgenic line with the inducibility of the reverse tetracycline transactivator (rtTA)/tetracycline-responsive element (tet-O)-driven transgenes. To ensure reliable rtTA expression in a broad range of cell types, we have targeted the rtTA transgene into the ROSA26 locus. The rtTA expression, however, is conditional to a Cre recombinase-mediated excision of a STOP region from the ROSA26 locus. We demonstrate the utility of this technology through the inducible expression of the vascular endothelial growth factor (VEGF-A) during embryonic development and postnatally in adult mice. Our results of adult induction recapitulate several different hepatic and immune cell pathological phenotypes associated with increased systemic VEGF-A protein levels. This system will be useful for studying genes in which temporal control of expression is necessary for the discovery of the full spectrum of functions. The presented approach abrogates the need to generate tissue-specific rtTA transgenes for tissues where well-characterized Cre lines already exist.

Present addresses: Gusztav Belteki, Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SA, UK

Jody Haigh and Katharina Haigh, Department for Molecular Biomedical Research, Flanders Interuniversity Institute of Biotechnology (VIB), Ghent University, Technologiepark 927, B-9052 Ghent (Zwijnaarde), Belgium

The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors


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