A limited number of double-strand DNA breaks is sufficient to delay cell cycle progression

Jeroen van den Berg, Anna G. Manjón, Karoline Kielbassa, Femke M. Feringa, Raimundo Freire, René H. Medema*

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

DNA damaging agents cause a variety of lesions, of which DNA double-strand breaks (DSBs) are the most genotoxic. Unbiased approaches aimed at investigating the relationship between the number of DSBs and outcome of the DNA damage response have been challenging due to the random nature in which damage is induced by classical DNA damaging agents. Here, we describe a CRISPR/Cas9-based system that permits us to efficiently introduce DSBs at defined sites in the genome. Using this system, we show that a guide RNA targeting only a single site in the human genome can trigger a checkpoint response that is potent enough to delay cell cycle progression. Abrogation of this checkpoint leads to DNA breaks in mitosis which gives rise to aneuploid progeny.

Original languageEnglish
Pages (from-to)10132-10144
Number of pages13
JournalNucleic acids research
Volume46
Issue number19
DOIs
Publication statusPublished - 1 Jan 2018
Externally publishedYes

Funding

The Dutch Cancer Foundation (KWF) [NKI 2014-6787]; Netherlands Organization for Scientific Research (NWO) [022.001.003]; Top-Go ZonMw [91210065]. Funding for open access charge: Top-Go ZonMw [91210065]. Conflict of interest statement. None declared. We thank Bas van Steensel, Eva Brinkman, Joppe Nieuwen-huis (Netherlands Cancer Institute) and Aniek Janssen (Lawrence Berkeley National Laboratory) for advice and critical reading of the manuscript. We thank all Medema, Rowland and Jacobs lab members for the helpful discussions on this study. The Dutch Cancer Foundation (KWF) [NKI 2014-6787]; Netherlands Organization for Scientific Research (NWO) [022.001.003]; Top-Go ZonMw [91210065]. Funding for open access charge: Top-Go ZonMw [91210065]. Conflict of interest statement. None declared.

FundersFunder number
Dutch Cancer Foundation
Netherlands Organization for Scientific Research
Lawrence Berkeley National Laboratory
ZonMw91210065
Nederlandse Organisatie voor Wetenschappelijk Onderzoek022.001.003
KWF KankerbestrijdingNKI 2014-6787
ICIN Netherlands Heart Institute

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