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Conditional cell ablation by tight control of caspase-3 dimerization in transgenic mice

Nature Biotechnology volume 20, pages 1234–1239 (2002)Cite this article

Abstract

Studying the effects of the loss of a specific cell type is a powerful approach in biology. Here we present a method based on the controlled activation of the apoptotic machinery. We expressed a modified caspase-3-containing chemical inducer of dimerization (CID)-binding sites in the livers of transgenic mice. In the absence of CID, no liver injury was detectable, underlining the absence of leakage in our system. In contrast, injection of the CID produced activation of the chimeric caspase-3, which led to a dose-dependent pure hepatocyte ablation with subsequent regeneration. This method is effective in both growing and nongrowing cells, and is therefore applicable to a wide range of cells and tissues. Moreover, because apoptosis has been described in numerous pathological circumstances, this system is useful for generating mouse models of human disorders as well as for studying the recovery or regeneration of tissues after cell loss.

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Figure 1: Construction, mechanism of caspase-3 activation, and analysis of transgenic animals.
Figure 2: Representative liver sections of transgenic animals.
Figure 3: Kinetics of caspase-3 activation and hepatocyte cytolysis in transgenic animals.
Figure 4: Kinetics of liver damage and regeneration in transgenic animals.
Figure 5: Dose dependence and CID re-injection.
Figure 6: Cytochrome c release in the liver of CID-injected mice.

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Acknowledgements

We thank Terry Van Dyke for the TTR vector and ARIAD Pharmaceuticals for providing us with AP20187 (http://www.ariad.com/regulationkits). We also thank Eva Mezey for proofreading the manuscript.

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Author notes

  1. Vincent O. Mallet and Claudia Mitchell: These two authors contributed equally to this work.

Authors and Affiliations

  1. Department of Genetics, Development and Molecular Pathology, Cochin Institute, 24 rue du Faubourg Saint Jacques, Paris, 75014, France

    Vincent O. Mallet, Claudia Mitchell, Jacques-Emmanuel Guidotti, Axel Kahn & Hélène Gilgenkrantz

  2. Laboratory of Biochemistry, Cochin Hospital, 27 rue du Faubourg Saint Jacques, Paris, 75014, France

    Patrick Jaffray

  3. Laboratory of Pathology, Bicêtre Hospital, 78 rue du Général Leclerc, Le Kremlin Bicêtre, 94275, France

    Monique Fabre

  4. Department of Immunology, Baylor College of Medicine, Houston, 77030, TX

    David Spencer

  5. EMI-U 9922 INSERM, Bichat-Claude Bernard Hospital, 46, rue Henri Huchard, Paris, 75877 cedex 18, France

    Damien Arnoult

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  1. Vincent O. Mallet
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Correspondence to Hélène Gilgenkrantz.

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Mallet, V., Mitchell, C., Guidotti, JE. et al. Conditional cell ablation by tight control of caspase-3 dimerization in transgenic mice. Nat Biotechnol 20, 1234–1239 (2002). https://doi.org/10.1038/nbt762

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