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Homing Endonuclease Target Determination Using SELEX Adapted for Yeast Surface Display

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Homing Endonucleases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1123))

Abstract

Knowing the target sequence of a DNA-binding protein is vital in obtaining fundamental characteristics of the protein and evaluating properties of the protein–DNA interaction. For example, novel homing endonucleases cannot be proven to be functional until a predicted target site is tested. Unfortunately, target site prediction is not always easy, or even possible, depending on the amount of sequence data available. Here we describe a modification of SELEX using yeast surface display that can quickly and inexpensively resolve DNA-binding targets in high throughput for proteins without any prior assumptions or knowledge regarding the target site. This protocol is easily integrated into the yeast surface display pipeline and is leveraged by the expansive number of existing tools for both SELEX and yeast surface display.

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Author information

Authors and Affiliations

  1. Program in Molecular and Cellular Biology and Department of Immunology, University of Washington, Seattle, WA, USA

    Kyle Jacoby & Andrew M. Scharenberg

  2. Center of Immunity and Immunotherapies, Seattle Children’s Research Institute, Seattle, WA, USA

    Kyle Jacoby & Andrew M. Scharenberg

  3. Northwest Genome Engineering Consortium, Seattle, WA, USA

    Andrew M. Scharenberg

Authors
  1. Kyle Jacoby
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  2. Andrew M. Scharenberg
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Editor information

Editors and Affiliations

  1. University of Western Ontario, London, Ontario, Canada

    David R. Edgell

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© 2014 Springer Science+Business Media New York

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Jacoby, K., Scharenberg, A.M. (2014). Homing Endonuclease Target Determination Using SELEX Adapted for Yeast Surface Display. In: Edgell, D. (eds) Homing Endonucleases. Methods in Molecular Biology, vol 1123. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-968-0_13

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  • DOI: https://doi.org/10.1007/978-1-62703-968-0_13

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-967-3

  • Online ISBN: 978-1-62703-968-0

  • eBook Packages: Springer Protocols

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