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

View ORCID ProfileJeroen van den Berg, Anna G. Manjón, Karoline Kielbassa, Femke M. Feringa, View ORCID ProfileRaimundo Freire, View ORCID ProfileRené H. Medema
doi: https://doi.org/10.1101/316158
Jeroen van den Berg
1Oncode Institute, Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands
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Anna G. Manjón
1Oncode Institute, Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands
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Karoline Kielbassa
1Oncode Institute, Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands
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Femke M. Feringa
1Oncode Institute, Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands
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Raimundo Freire
2Unidad de Investigación, Hospital Universitario de Canarias, Instituto de Tecnologías Biomédicas, Ofra s/n, La Laguna, Tenerife, Spain
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René H. Medema
1Oncode Institute, Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands
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  • For correspondence: r.medema@nki.nl
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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 give rise to micronucleated daughter cells.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted May 07, 2018.
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A limited number of double-strand DNA breaks are sufficient to delay cell cycle progression
Jeroen van den Berg, Anna G. Manjón, Karoline Kielbassa, Femke M. Feringa, Raimundo Freire, René H. Medema
bioRxiv 316158; doi: https://doi.org/10.1101/316158
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A limited number of double-strand DNA breaks are sufficient to delay cell cycle progression
Jeroen van den Berg, Anna G. Manjón, Karoline Kielbassa, Femke M. Feringa, Raimundo Freire, René H. Medema
bioRxiv 316158; doi: https://doi.org/10.1101/316158

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