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Targeted Gene Knock Out Using Nuclease-Assisted Vector Integration: Hemi- and Homozygous Deletion of JAG1

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Synthetic Biology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1772))

Abstract

Gene editing technologies are revolutionizing fields such as biomedicine and biotechnology by providing a simple means to manipulate the genetic makeup of essentially any organism. Gene editing tools function by introducing double-stranded breaks at targeted sites within the genome, which the host cells repair preferentially by Non-Homologous End Joining. While the technologies to introduce double-stranded breaks have been extensively optimized, this progress has not been matched by the development of methods to integrate heterologous DNA at the target sites or techniques to detect and isolate cells that harbor the desired modification. We present here a technique for rapid introduction of vectors at target sites in the genome that enables efficient isolation of successfully edited cells.

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Author information

Authors and Affiliations

  1. Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL, USA

    Michael Gapinske, Nathan Tague, Jackson Winter, Gregory H. Underhill & Pablo Perez-Pinera

  2. Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USA

    Gregory H. Underhill & Pablo Perez-Pinera

Authors
  1. Michael Gapinske
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  2. Nathan Tague
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  3. Jackson Winter
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  4. Gregory H. Underhill
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  5. Pablo Perez-Pinera
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Corresponding author

Correspondence to Pablo Perez-Pinera .

Editor information

Editors and Affiliations

  1. Agilent Technologies, Inc, La Jolla, California, USA

    Jeffrey Carl Braman

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Gapinske, M., Tague, N., Winter, J., Underhill, G.H., Perez-Pinera, P. (2018). Targeted Gene Knock Out Using Nuclease-Assisted Vector Integration: Hemi- and Homozygous Deletion of JAG1. In: Braman, J. (eds) Synthetic Biology. Methods in Molecular Biology, vol 1772. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7795-6_13

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  • DOI: https://doi.org/10.1007/978-1-4939-7795-6_13

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7794-9

  • Online ISBN: 978-1-4939-7795-6

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