DNA damage response (DDR) is critically important for cell survival, genome maintenance, and its defect has been exploited therapeutically in cancer treatment. Many DDR-targeting agents have been generated and have entered the clinic and/or clinical trials. In order to provide a global and unbiased view of DDR network, we designed a focused CRISPR library targeting 365 DDR genes and performed CRISPR screens on the responses to several DDR inhibitors and DNA-damaging agents in 293A cells. With these screens, we determined responsive pathways enriched under treatment with different types of small-molecule agents. Additionally, we showed that POLE3/4-deficient cells displayed enhanced sensitivity to an ATR inhibitor, a PARP inhibitor, and camptothecin. Moreover, by performing DDR screens in isogenic TP53 wild-type and TP53 knock-out cell lines, our results suggest that the performance of our CRISPR DDR dropout screens is independent of TP53 status. Collectively, our findings indicate that CRISPR DDR screens can be used to identify potential targets of small-molecule drugs and reveal that TP53 status does not affect the outcome of these screens.

中文翻译：

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基于CRISPR / CAS9的DNA损伤反应筛选揭示了基因-药物相互作用。

DNA损伤反应（DDR）对于细胞存活，基因组维护至关重要，并且其缺陷已在癌症治疗中得到治疗性利用。已经产生了许多DDR靶向剂，并已进入临床和/或临床试验。为了提供DDR网络的全球公正视图，我们设计了针对365个DDR基因的聚焦CRISPR文库，并对293A细胞对几种DDR抑制剂和DNA损伤剂的反应进行了CRISPR筛选。通过这些筛选，我们确定了在不同类型的小分子药物治疗下富集的反应途径。此外，我们显示POLE3 / 4缺陷细胞对ATR抑制剂，PARP抑制剂和喜树碱的敏感性增强。此外，通过在同基因TP53野生型和TP53敲除细胞系中进行DDR筛选，我们的结果表明，我们的CRISPR DDR辍学筛选的性能与TP53状态无关。总的来说，我们的发现表明，CRISPR DDR筛选可用于识别小分子药物的潜在靶标，并揭示TP53的状态不会影响这些筛选的结果。