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Conversion Tract Analysis of Homology-Directed Genome Editing Using Oligonucleotide Donors

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DNA Repair

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1999))

Abstract

Homology-directed genome editing is the intentional alteration of an endogenous genetic locus using information from an exogenous homology donor. A conversion tract is defined as the amount of genetic information that is converted from the homology donor to a given strand of the targeted chromosomal locus. Because of this, conversion tract analysis retrospectively not only elucidates the mechanism of homology-directed genome editing but also provides valuable insights on the conversion efficiency of every nucleotide in the homology donor. Here we describe a blue fluorescent protein-to-green fluorescent protein conversion system that can be conveniently used to measure the efficiency and analyze the lengths of conversion tracts of homology-directed genome editing using oligonucleotide donors in mammalian cells.

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Author information

Authors and Affiliations

  1. eGenesis, Cambridge, MA, USA

    Yinan Kan

  2. BMBB Department, University of Minnesota Medical School, Minneapolis, MN, USA

    Eric A. Hendrickson

Authors
  1. Yinan Kan
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  2. Eric A. Hendrickson
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Corresponding author

Correspondence to Eric A. Hendrickson .

Editor information

Editors and Affiliations

  1. Indiana University – Purdue University, Indianapolis, IN, USA

    Lata Balakrishnan

  2. Department of Biological Sciences, University of South Carolina, Columbia, SC, USA

    Jason A. Stewart

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Kan, Y., Hendrickson, E.A. (2019). Conversion Tract Analysis of Homology-Directed Genome Editing Using Oligonucleotide Donors. In: Balakrishnan, L., Stewart, J. (eds) DNA Repair. Methods in Molecular Biology, vol 1999. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9500-4_7

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  • DOI: https://doi.org/10.1007/978-1-4939-9500-4_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9499-1

  • Online ISBN: 978-1-4939-9500-4

  • eBook Packages: Springer Protocols

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