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Genome Editing in Human Pluripotent Stem Cells

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Stem Cell Banking

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1590))

Abstract

Genome editing in human pluripotent stem cells (hPSCs) enables the generation of reporter lines and knockout cell lines. Zinc finger nucleases, transcription activator-like effector nucleases (TALENs), and CRISPR/Cas9 technology have recently increased the efficiency of proper gene editing by creating double strand breaks (DSB) at defined sequences in the human genome. These systems typically use plasmids to transiently transcribe nucleases within the cell. Here, we describe the process for preparing hPSCs for transient expression of nucleases via electroporation and subsequent analysis to create genetically modified stem cell lines.

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Author information

Authors and Affiliations

  1. Department of Biomedical Engineering, University of Wisconsin–Madison, 1550 Engineering Drive, Madison, WI, 53706, USA

    Jared Carlson-Stevermer & Krishanu Saha

  2. Wisconsin Institute for Discovery, University of Wisconsin–Madison, 330 N. Orchard Street, Madison, WI, 53715, USA

    Jared Carlson-Stevermer & Krishanu Saha

Authors
  1. Jared Carlson-Stevermer
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  2. Krishanu Saha
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Corresponding author

Correspondence to Krishanu Saha .

Editor information

Editors and Affiliations

  1. ARC Centre of Excellence for Electromaterials Science, Intelligent Polymer Research Institute, AIIM Facility, Innovation Campus, University of Wollongong, Squires Way, Fairy Meadow, New South Wales, Australia

    Jeremy M. Crook

  2. WiCell Research Institute, Madison, Wisconsin, USA

    Tenneille E. Ludwig

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Carlson-Stevermer, J., Saha, K. (2017). Genome Editing in Human Pluripotent Stem Cells. In: Crook, J., Ludwig, T. (eds) Stem Cell Banking. Methods in Molecular Biology, vol 1590. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6921-0_12

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  • DOI: https://doi.org/10.1007/978-1-4939-6921-0_12

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6919-7

  • Online ISBN: 978-1-4939-6921-0

  • eBook Packages: Springer Protocols

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