Enhancement of homology-directed repair with chromatin donor templates in cells

  1. Grisel Cruz-Becerra
  2. James T Kadonaga  Is a corresponding author
  1. University of California, San Diego, United States

Abstract

A key challenge in precise genome editing is the low efficiency of homology-directed repair (HDR). Here we describe a strategy for increasing the efficiency of HDR in cells by using a chromatin donor template instead of a naked DNA donor template. The use of chromatin, which is the natural form of DNA in the nucleus, increases the frequency of HDR-edited clones as well as homozygous editing. In addition, transfection of chromatin results in negligible cytotoxicity. These findings suggest that a chromatin donor template should be useful for a wide range of HDR applications such as the precise insertion or replacement of DNA fragments that contain the coding regions of genes.

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All data generated or analysed during this study are included in the manuscript and supporting files.

Article and author information

Author details

  1. Grisel Cruz-Becerra

    Section of Molecular Biology, University of California, San Diego, La Jolla, United States
    Competing interests
    Grisel Cruz-Becerra, has filed a patent application (PCT/US2019/029194) that describes the invention reported in this article.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6297-4132
  2. James T Kadonaga

    Section of Molecular Biology, University of California, San Diego, La Jolla, United States
    For correspondence
    jkadonaga@ucsd.edu
    Competing interests
    James T Kadonaga, has filed a patent application (PCT/US2019/029194) that describes the invention reported in this article.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2075-9458

Funding

National Institutes of Health (R35 GM118060)

  • James T Kadonaga

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2020, Cruz-Becerra & Kadonaga

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Grisel Cruz-Becerra
  2. James T Kadonaga
(2020)
Enhancement of homology-directed repair with chromatin donor templates in cells
eLife 9:e55780.
https://doi.org/10.7554/eLife.55780

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https://doi.org/10.7554/eLife.55780

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