A limited number of double-strand DNA breaks is sufficient to delay cell cycle progression
A limited number of double-strand DNA breaks is sufficient to delay cell cycle progression
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DOI：
10.1093/nar/gky786
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发表时间：
2018-11-02
影响因子：
14.9
通讯作者：
Medema, Rene H.
中科院分区：
文献类型：
--
作者：
van den Berg, Jeroen;Manjon, Anna G.;Medema, Rene H.
DNA损伤剂可引起多种损伤，其中DNA双链断裂(DSB)具有最强的遗传毒性。由于经典DNA损伤剂引起损伤的随机性，旨在研究DSB数量和DNA损伤反应结果之间关系的无偏见方法一直是具有挑战性的。在这里，我们描述了一个基于CRISPR/CAS9的系统，它允许我们在基因组中定义的位置高效地引入DSB。使用这个系统，我们证明了只针对人类基因组中的一个单一位置的引导RNA可以触发足以延缓细胞周期进展的检查点反应。取消这一检查点会导致有丝分裂中的DNA断裂，从而产生非整倍体后代。
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.