Molecular safeguarding of CRISPR gene drive experiments

Abstract

CRISPR-based homing gene drives have sparked both enthusiasm and deep concerns due to their potential for genetically altering entire species. This raises the question about our ability to prevent the unintended spread of such drives from the laboratory into a natural population. Here, we experimentally demonstrate the suitability of synthetic target site drives as well as split drives as flexible safeguarding strategies for gene drive experiments by showing that their performance closely resembles that of standard homing drives in Drosophila melanogaster. Using our split drive system, we further find that maternal deposition of both Cas9 and gRNA is required to form resistance alleles in the early embryo and that maternally-deposited Cas9 alone can power germline drive conversion in individuals that lack a genomic source of Cas9.

Data availability

All data generated are available in Supplementary file 2.

Article and author information

Author details

  1. Jackson Champer

    Department of Biological Statistics and Computational Biology, Cornell University, Ithaca, United States
    For correspondence
    jc3248@cornell.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3814-3774
  2. Joan Chung

    Department of Biological Statistics and Computational Biology, Cornell University, Ithaca, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Yoo Lim Lee

    Department of Biological Statistics and Computational Biology, Cornell University, Ithaca, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Chen Liu

    Department of Biological Statistics and Computational Biology, Cornell University, Ithaca, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Emily Yang

    Department of Biological Statistics and Computational Biology, Cornell University, Ithaca, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Zhaoxin Wen

    Department of Biological Statistics and Computational Biology, Cornell University, Ithaca, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Andrew G Clark

    Department of Biological Statistics and Computational Biology, Cornell University, Ithaca, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Philipp W Messer

    Department of Biological Statistics and Computational Biology, Cornell University, Ithaca, United States
    For correspondence
    messer@cornell.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8453-9377

Funding

National Institutes of Health (R21AI130635)

  • Jackson Champer
  • Andrew G Clark
  • Philipp W Messer

National Institutes of Health (F32AI138476)

  • Jackson Champer

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

Reviewing Editor

  1. Kevin M Esvelt, Massachusetts Institute of Technology, United States

Publication history

  1. Received: August 25, 2018
  2. Accepted: January 9, 2019
  3. Accepted Manuscript published: January 22, 2019 (version 1)
  4. Version of Record published: February 1, 2019 (version 2)

Copyright

© 2019, Champer et al.

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. Jackson Champer
  2. Joan Chung
  3. Yoo Lim Lee
  4. Chen Liu
  5. Emily Yang
  6. Zhaoxin Wen
  7. Andrew G Clark
  8. Philipp W Messer
(2019)
Molecular safeguarding of CRISPR gene drive experiments
eLife 8:e41439.
https://doi.org/10.7554/eLife.41439

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