Article | Published:

Safeguarding CRISPR-Cas9 gene drives in yeast

Nature Biotechnology volume 33, pages 12501255 (2015) | Download Citation

Abstract

RNA-guided gene drives capable of spreading genomic alterations made in laboratory organisms through wild populations could be used to address environmental and public health problems. However, the possibility of unintended genome editing occurring through the escape of strains from laboratories, coupled with the prospect of unanticipated ecological change, demands caution. We report the efficacy of CRISPR-Cas9 gene drive systems in wild and laboratory strains of the yeast Saccharomyces cerevisiae. Furthermore, we address concerns surrounding accidental genome editing by developing and validating methods of molecular confinement that minimize the risk of unwanted genome editing. We also present a drive system capable of overwriting the changes introduced by an earlier gene drive. These molecular safeguards should enable the development of safe CRISPR gene drives for diverse organisms.

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Acknowledgements

We are very grateful to S. Doris, D. Spatt and F. Winston for their incredible patience, generosity and expertise in tetrad dissection. We also thank F. Winston for providing us with SK1 strains and members of the Church laboratory for insightful discussions. This work was supported by grants from the Department of Energy (DOE) (DE-FG02-02ER63445 to G.M.C.), National Science Foundation (NSF) (SynBERC SA5283-11210 and MCB-1330914 to G.M.C.), National Cancer Institute (NCI) (5T32CA009216-34 to A.C.), National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) (1K99DK102669-01 to K.M.E.) and the Wyss Institute for Biologically Inspired Engineering (Technology Development Fellowship to K.M.E.).

Author information

Author notes

    • James E DiCarlo
    •  & Alejandro Chavez

    These authors contributed equally to this work.

Affiliations

  1. Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.

    • James E DiCarlo
    • , Alejandro Chavez
    • , Sven L Dietz
    •  & George M Church
  2. Harvard Medical School, Boston, Massachusetts, USA.

    • James E DiCarlo
    • , Alejandro Chavez
    • , Sven L Dietz
    • , Kevin M Esvelt
    •  & George M Church
  3. Department of Biomedical Engineering, Boston University, Boston, Massachusetts, USA.

    • James E DiCarlo
  4. Wyss Institute for Biologically Inspired Engineering, Boston, Massachusetts, USA.

    • Alejandro Chavez
    • , Sven L Dietz
    • , Kevin M Esvelt
    •  & George M Church
  5. Department of Pathology, Massachusetts General Hospital, Boston, Massachusetts, USA.

    • Alejandro Chavez
  6. Department for Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland.

    • Sven L Dietz

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Contributions

S.L.D. initiated the study; J.E.D., A.C., S.L.D. and K.M.E. designed the experiments; J.E.D. performed the experiments with assistance from A.C.; J.E.D., A.C., S.L.D. and K.M.E. analyzed the data; and K.M.E. wrote the paper with A.C. and contributing input from J.E.D., S.L.D. and G.M.C.

Competing interests

K.M.E. and G.M.C. are authors of a patent filed on CRISPR gene drive (PCT/US2015/010550). K.M.E. is author of a provisional patent filed on CRISPR gene drive (serial no. 62/236,545).

Corresponding authors

Correspondence to Kevin M Esvelt or George M Church.

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DOI

https://doi.org/10.1038/nbt.3412

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