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Somatic Cell Nuclear Transfer in the Mouse

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Microinjection

Part of the book series: Methods in Molecular Biology ((MIMB,volume 518))

Abstract

Somatic cell nuclear transfer (SCNT) has become a unique and powerful tool for epigenetic reprogramming research and gene manipulation in animals since “Dolly,” the first animal cloned from an adult cell was reported in 1997. Although the success rates of somatic cloning have been inefficient and the mechanism of reprogramming is still largely unknown, this technique has been proven to work in more than 10 mammalian species. Among them, the mouse provides the best model for both basic and applied research of somatic cloning because of its abounding genetic resources, rapid sexual maturity and propagation, minimal requirements for housing, etc. This chapter describes a basic protocol for mouse cloning using cumulus cells, the most popular cell type for NT, in which donor nuclei are directly injected into the oocyte using a piezo-actuated micromanipulator. In particular, we focus on a new, more efficient mouse cloning protocol using trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, which increases both in vitro and in vivo developmental rates from twofold to fivefold. This new method including TSA will be helpful to establish mouse cloning in many laboratories.

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Acknowledgments

We thank Dr. T. Castranio and all laboratory members for discussion and critical reading of the manuscript. This work in our laboratory was supported by RIKEN (Strategic Program for Research and Development (FY2005) to S.K.) and the Ministry of Education, Culture, Sports, Science and Technology of Japan (17780213 to S.K., 15080211 to T.W., and the Project for the Realization of Regenerative Medicine to T.W.)

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Authors and Affiliations

  1. RIKEN, Center for Developmental Biology, Kobe, Japan

    Satoshi Kishigami & Teruhiko Wakayama

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  1. Satoshi Kishigami
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  2. Teruhiko Wakayama
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Editors and Affiliations

  1. Dept. Biological Sciences, Florida Institute of Technology, W. University Boulevard 150, Melbourne, 32901, U.S.A.

    David J. Carroll

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Kishigami, S., Wakayama, T. (2009). Somatic Cell Nuclear Transfer in the Mouse. In: Carroll, D. (eds) Microinjection. Methods in Molecular Biology, vol 518. Humana Press. https://doi.org/10.1007/978-1-59745-202-1_15

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  • DOI: https://doi.org/10.1007/978-1-59745-202-1_15

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-884-3

  • Online ISBN: 978-1-59745-202-1

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