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Induced Pluripotent Stem (iPS) Cells pp 111–131Cite as

Inducible Transgene Expression in Human iPS Cells Using Versatile All-in-One piggyBac Transposons

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1357))

Abstract

Transgenics is a mainstay of functional genomics. Conditionally overexpressing genes of interest (GOIs) helps to reveal their roles in the control of complex biological processes. Complemented by findings in classic animal model systems, recent advances in human embryonic stem cell (hESC) and patient-specific induced pluripotent stem cell (hiPSC) differentiation have led to sophisticated in vitro models of human development and disease. Yet, as transgenic elements encoding inducible systems must be introduced de novo into each genetically unique human stem cell line, robust and straightforward solutions to gene delivery are required. Transposons are a family of mobile DNA elements that have been adapted as experimental tools for stable genomic integration of transgenes. The piggyBac (PB) transposon from Trichoplusia ni presents a number of benefits over classic viral or BAC transgenesis: ease of application, simple integration-site mapping, and the unique capacity for traceless excision. Moreover, their large capacity permits the consolidation of multiple transgene components in a single vector system. In this chapter, we outline the features of a panel of “All-in-One” PB transposons designed for drug-inducible gene expression and provide guidelines to establish and validate populations or clones of transgenic hiPSCs.

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Acknowledgments

We like to recognize Ryoko Hirohata and Tomoko Matsumoto for their valuable technical support in developing these materials and protocols. Our appreciation to all our colleagues who continue to make use of these transposon resources, in various research applications. This work was supported by the Research Center Network for Realization of Regenerative Medicine, Program for Intractable Diseases Research Utilizing Disease-Specific iPS Cells, of the Japan Science and Technology Agency (JST), and the Strategic International Collaborative Research Program of the JST. K.W. is a Hakubi Center Special Project Researcher. S.Y. is a non-salaried scientific advisor of iPS Academia Japan. S-I.K. is a JSPS Fellowship recipient (2011–2013) and JST Researcher.

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Authors and Affiliations

  1. Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, 606-8507, Japan

    Shin-Il Kim, Fabian Oceguera-Yanez, Chiho Sakurai, Masato Nakagawa, Shinya Yamanaka & Knut Woltjen

  2. Gladstone Institutes of Cardiovascular Disease, University of California, San Francisco, CA, 94158, USA

    Shinya Yamanaka

  3. Hakubi Center for Advanced Research, Kyoto University, Kyoto, 606-8501, Japan

    Knut Woltjen

Authors
  1. Shin-Il Kim
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  2. Fabian Oceguera-Yanez
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  3. Chiho Sakurai
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  4. Masato Nakagawa
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  5. Shinya Yamanaka
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  6. Knut Woltjen
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Corresponding author

Correspondence to Knut Woltjen .

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Editors and Affiliations

  1. Ottawa Hospital Research Institute, Ottawa, Ontario, Canada

    Kursad Turksen

  2. Mount Sinai Hospital Samuel Lunenfeld Research Inst, Toronto, Ontario, Canada

    Andras Nagy

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Kim, SI., Oceguera-Yanez, F., Sakurai, C., Nakagawa, M., Yamanaka, S., Woltjen, K. (2015). Inducible Transgene Expression in Human iPS Cells Using Versatile All-in-One piggyBac Transposons. In: Turksen, K., Nagy, A. (eds) Induced Pluripotent Stem (iPS) Cells. Methods in Molecular Biology, vol 1357. Humana Press, New York, NY. https://doi.org/10.1007/7651_2015_251

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  • DOI: https://doi.org/10.1007/7651_2015_251

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3054-8

  • Online ISBN: 978-1-4939-3055-5

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