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Generation of PDE4 Knockout Mice by Gene Targeting

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Phosphodiesterase Methods and Protocols

Part of the book series: Methods In Molecular Biology™ ((MIMB,volume 307))

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Abstract

The development of gene-targeting techniques has ushered in a new era in mouse genetics. Two discoveries have been instrumental: the finding that an exogenous DNA introduced in mammalian cells can recombine with homologous chromosomal sequences, a process known as gene targeting, and the revelation that cultured embryonic stem (ES) cells when injected into early stage mouse embryos can contribute to produce germ-line chimeras. On the basis of these seminal findings, gene targeting by homologous recombination in mouse ES cells in vitro has been established as a powerful means of altering specific loci in the mouse genome. As a result, gene function can be studied in vivo. By applying this technology, targeted disruption of PDE4 alleles is created in cultured ES cells and, subsequently, the mutant ES cells are injected into blastocysts and returned to pseudopregnant foster mothers to produce germ-line chimeric pups. In this chapter, we describe the basic protocols used to generate the PDE4 knockout mice.

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Acknowledgments

We are indebted to Dr. Beverly Koller for assistance with the technical details described herein. This work was supported by National Institutes of Health grants HD20788, HL67674, and HD5344.

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Authors and Affiliations

  1. Division of Reproductive Biology, Department of Obstetrics and Gynecology, Stanford University Medical Center Stanford, CA

    S.-L. Catherine Jin & Marco Conti

  2. Department of Medicine, University of North Carolina, Chapel Hill, NC

    Anne M. Latour

Authors
  1. S.-L. Catherine Jin
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  2. Anne M. Latour
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  3. Marco Conti
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Editors and Affiliations

  1. Pharmacolgoie et Physico-Chimie des Interactions Celluaires et Moléculaires, Centre National de la Recherche Scientifique, Université Louis Pasteur de Strasbourg, Faculté de Pharmacie, Illkirch, France

    Claire Lugnier

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© 2005 Humana Press Inc.

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Jin, SL.C., Latour, A.M., Conti, M. (2005). Generation of PDE4 Knockout Mice by Gene Targeting. In: Lugnier, C. (eds) Phosphodiesterase Methods and Protocols. Methods In Molecular Biology™, vol 307. Humana Press. https://doi.org/10.1385/1-59259-839-0:191

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  • DOI: https://doi.org/10.1385/1-59259-839-0:191

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-314-5

  • Online ISBN: 978-1-59259-839-7

  • eBook Packages: Springer Protocols

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