Brief Communication | Published:

CRISPR–Cas9 genome editing induces a p53-mediated DNA damage response

Nature Medicine (2018) | Download Citation

Abstract

Here, we report that genome editing by CRISPR–Cas9 induces a p53-mediated DNA damage response and cell cycle arrest in immortalized human retinal pigment epithelial cells, leading to a selection against cells with a functional p53 pathway. Inhibition of p53 prevents the damage response and increases the rate of homologous recombination from a donor template. These results suggest that p53 inhibition may improve the efficiency of genome editing of untransformed cells and that p53 function should be monitored when developing cell-based therapies utilizing CRISPR–Cas9.

  • Subscribe to Nature Medicine for full access:

    $225

    Subscribe

Additional access options:

Already a subscriber?  Log in  now or  Register  for online access.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

References

  1. 1.

    Hustedt, N. & Durocher, D. Nat. Cell Biol. 19, 1–9 (2016).

  2. 2.

    Hohmann, S. & Gozalbo, D. Mol. Gen. Genet. 211, 446–454 (1988).

  3. 3.

    Richardson, C. D., Ray, G. J., DeWitt, M. A., Curie, G. L. & Corn, J. E. Nat. Biotechnol. 34, 339–344 (2016).

  4. 4.

    DeWitt, M. A. et al. Sci. Transl. Med. 8, 360ra134 (2016).

  5. 5.

    Yin, H. et al. Nat. Biotechnol. 32, 551–553 (2014).

  6. 6.

    Dever, D. P. et al. Nature 539, 384–389 (2016).

  7. 7.

    Lee, K. et al. eLife 6, e25312 (2017).

  8. 8.

    Maruyama, T. et al. Nat. Biotechnol. 33, 538–542 (2015).

  9. 9.

    Schmierer, B. et al. Mol. Syst. Biol. 13, 945 (2017).

  10. 10.

    Luo, M. & Chen, Y. Int. J. Ophthalmol. 11, 150–159 (2018).

  11. 11.

    Otto, T. & Sicinski, P. Nat. Rev. Cancer 17, 93–115 (2017).

  12. 12.

    Sokolova, M. et al. Cell Cycle 16, 189–199 (2017).

  13. 13.

    Doench, J. G. et al. Nat. Biotechnol. 34, 184–191 (2016).

  14. 14.

    Wang, J., Vasaikar, S., Shi, Z., Greer, M. & Zhang, B. Nucleic Acids Res. 45, W130–W137 (2017).

  15. 15.

    Canny, M. D. et al. Nat. Biotechnol. 36, 95–102 (2018).

  16. 16.

    Cuella-Martin, R. et al. Mol. Cell 64, 51–64 (2016).

  17. 17.

    Muerdter, F. et al. Nat. Methods 15, 141–149 (2018).

  18. 18.

    Li, W. et al. Genome Biol. 15, 554 (2014).

  19. 19.

    Wang, T. et al. Science 350, 1096–1101 (2015).

  20. 20.

    Tsai, S. Q. et al. Nat. Biotechnol. 33, 187–197 (2015).

Download references

Acknowledgements

Part of this work was carried out at the High Throughput Genome Engineering Facility and the Swedish National Genomics Infrastructure funded by Science for Life Laboratory (Scilifelab). The Knut and Alice Wallenberg Foundation, Cancerfonden, Barncancerfonden and the Academy of Finland supported this work. We thank H. Han and Y. Bryceson for providing equipment, the Protein Science Facility at Karolinska Institutet, as well as I. Sur and T. Kivioja for their comments on the manuscript.

Author information

Author notes

  1. These authors contributed equally: Emma Haapaniemi, Sandeep Botla.

  2. These authors jointly supervised this work: Bernhard Schmierer, Jussi Taipale.

Affiliations

  1. Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden

    • Emma Haapaniemi
    • , Sandeep Botla
    • , Jenna Persson
    • , Bernhard Schmierer
    •  & Jussi Taipale
  2. Genome-Scale Biology Program, University of Helsinki, Helsinki, Finland

    • Emma Haapaniemi
    •  & Jussi Taipale
  3. Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom

    • Jussi Taipale

Authors

  1. Search for Emma Haapaniemi in:

  2. Search for Sandeep Botla in:

  3. Search for Jenna Persson in:

  4. Search for Bernhard Schmierer in:

  5. Search for Jussi Taipale in:

Contributions

E.H., B.S. and J.T. wrote the manuscript. S.B., B.S. and J.P. conducted the genome-wide knockout screens. E.H., B.S. and S.B. prepared the cell lines and performed the flow cytometry experiments. J.T. and B.S. supervised the study. All authors read and approved the final manuscript.

Competing interests

The authors declare no competing interests.

Corresponding authors

Correspondence to Bernhard Schmierer or Jussi Taipale.

Supplementary information

  1. Supplementary Text and Figures

    Supplementary Figures 1–5 and Supplementary Tables 1–3

  2. Reporting Summary

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/s41591-018-0049-z

Rights and permissions

To obtain permission to re-use content from this article visit RightsLink.