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Genetic Inducible Fate Mapping in Adult Mice Using Tamoxifen-Dependent Cre Recombinases

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Mouse Genetics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1194))

Abstract

The Cre/lox site-specific recombination system allows the control of gene activity in space and time in almost any tissue of the mouse. A major technical advance was the development of tamoxifen-dependent Cre recombinases, such as CreERT2, that can be activated by administration of tamoxifen to the animal. This powerful tool greatly facilitates the study of gene functions and the generation of more realistic animal models of sporadic human diseases. Another important application of tamoxifen-dependent Cre recombinases is genetic inducible fate mapping (GIFM). In GIFM studies, the inducible Cre/lox system is used to genetically label a defined cell population at a selected time by irreversible activation of the expression of a Cre-responsive reporter transgene. Then, marked cells are detected at later time points to determine how the originally labeled progenitors contribute to specific structures and cell types during pre- and postnatal development. GIFM was initially applied during mouse embryogenesis, but is now increasingly used for cell lineage tracing in adult mice under physiological and pathophysiological conditions. Here we describe the design of GIFM experiments in adult mice as exemplified by CreERT2-assisted tracing of vascular smooth muscle cells during the development of atherosclerotic lesions. First, we give an overview of reporter transgenes available for genetic cell marking that are expressed from the Rosa26 locus, such as β-galactosidase and fluorescent proteins. Then we present detailed protocols for the generation of experimental mice for GIFM studies, the induction of cell labeling by tamoxifen treatment, and the detection of marked cells in fixed and live tissues. Each section also provides a discussion of limitations and common pitfalls of GIFM experiments. Most of the protocols can be easily adapted to other developmental stages, cell types, Cre recombinases, and reporter transgenes and, thus, can be used as general guidelines for GIFM studies in mice.

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Acknowledgments

We thank the members of the Feil laboratory for critical discussion. Work in the authors’ laboratory was supported by grants from VolkswagenStiftung, Deutsche Forschungsgemeinschaft, fortüne-Programm der Medizinischen Fakultät der Universität Tübingen, and Dr. Karl Kuhn-Stiftung.

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Authors and Affiliations

  1. Interfakultäres Institut für Biochemie, Universität Tübingen, Hoppe-Seyler-Str. 4, Tübingen, 72076, Germany

    Susanne Feil, Jana Krauss, Martin Thunemann & Robert Feil

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  1. Susanne Feil
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  2. Jana Krauss
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  3. Martin Thunemann
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  4. Robert Feil
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Corresponding author

Correspondence to Robert Feil .

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Editors and Affiliations

  1. Stem Cell Regulation and Animal Aging Section, Basic Research Laboratory,, National Cancer Institute, National Institutes of Health, Frederick, MD, USA

    Shree Ram Singh

  2. Genetically Engineered Mouse Modeling Core/Department of Molecular Virology, Immunology and Medical Genetics, Wexner Medical Center, Comprehensive Cancer Center, The Ohio State University, Columbus, OH, USA

    Vincenzo Coppola

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Feil, S., Krauss, J., Thunemann, M., Feil, R. (2014). Genetic Inducible Fate Mapping in Adult Mice Using Tamoxifen-Dependent Cre Recombinases. In: Singh, S., Coppola, V. (eds) Mouse Genetics. Methods in Molecular Biology, vol 1194. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1215-5_6

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  • DOI: https://doi.org/10.1007/978-1-4939-1215-5_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1214-8

  • Online ISBN: 978-1-4939-1215-5

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