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Engineering of Conditional Class I Hdac Knockout Mice and Generation of a Time-Spatial Knockout by a Dual Recombination System

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HDAC/HAT Function Assessment and Inhibitor Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1510))

Abstract

The protein sequences of class I HDACs in mice and humans are 96–99 % identical. These highly conserved proteins have crucial roles in biological processes, such as proliferation and development, which is reflected in the lethality that occurs in conventional whole body knockout mice. Therefore, conditional knockouts are inevitable to investigate the functions of class I HDACs in mice. Here, we describe the generation of conditional class I Hdac knockout mice, using Hdac1 as an example. We explain a relatively quick procedure to generate the necessary target vectors by recombination-mediated genetic engineering and gateway techniques. Furthermore, we show how to culture, target, and screen for positively recombined ES cells. Additionally, we present a dual recombination system, which allows the deletion of class I Hdacs at any time by a tamoxifen inducible Cre.

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Acknowledgment

This work was supported by Else Kröner Fresenius Stiftung (2016_A43) to MW.

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Authors and Affiliations

  1. Klinikum rechts der Isar, II. Medizinische Klinik, Technische Universität München, Ismaninger Str. 22, 81675, Munich, Germany

    Sieglinde Bayer & Matthias Wirth

Authors
  1. Sieglinde Bayer
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  2. Matthias Wirth
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Corresponding author

Correspondence to Matthias Wirth .

Editor information

Editors and Affiliations

  1. Institute of Toxicology, University Medical Center Mainz, Mainz, Germany

    Oliver H. Krämer

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Bayer, S., Wirth, M. (2017). Engineering of Conditional Class I Hdac Knockout Mice and Generation of a Time-Spatial Knockout by a Dual Recombination System. In: Krämer, O. (eds) HDAC/HAT Function Assessment and Inhibitor Development. Methods in Molecular Biology, vol 1510. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6527-4_14

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  • DOI: https://doi.org/10.1007/978-1-4939-6527-4_14

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6525-0

  • Online ISBN: 978-1-4939-6527-4

  • eBook Packages: Springer Protocols

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