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

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.

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Fig. 1: Unbiased CRISPR screening identifies activation of the p53–p21–RB axis.
Fig. 2: Cas9 gRNA RNP delivery triggers a p53-dependent DNA damage response that suppresses gene correction.

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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.

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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.

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Correspondence to Bernhard Schmierer or Jussi Taipale.

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Haapaniemi, E., Botla, S., Persson, J. et al. CRISPR–Cas9 genome editing induces a p53-mediated DNA damage response. Nat Med 24, 927–930 (2018). https://doi.org/10.1038/s41591-018-0049-z

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