The response to DNA damage is critical for cellular homeostasis, tumor suppression, immunity, and gametogenesis. In order to provide an unbiased and global view of the DNA damage response in human cells, we undertook 31 CRISPR-Cas9 screens against 27 genotoxic agents in the retinal pigment epithelium-1 (RPE1) cell line. These screens identified 890 genes whose loss causes either sensitivity or resistance to DNA-damaging agents. Mining this dataset, we discovered that ERCC6L2 (which is mutated in a bone-marrow failure syndrome) codes for a canonical non-homologous end-joining pathway factor, that the RNA polymerase II component ELOF1 modulates the response to transcription-blocking agents, and that the cytotoxicity of the G-quadruplex ligand pyridostatin involves trapping topoisomerase II on DNA. This map of the DNA damage response provides a rich resource to study this fundamental cellular system and has implications for the development and use of genotoxic agents in cancer therapy.

对 DNA 损伤的反应对于细胞稳态、肿瘤抑制、免疫和配子发生至关重要。为了对人类细胞中的 DNA 损伤反应提供公正和全面的看法，我们对视网膜色素上皮细胞 1 (RPE1) 细胞系中的 27 种基因毒性物质进行了 31 次 CRISPR-Cas9 筛选。这些筛选鉴定出 890 个基因，这些基因的缺失会导致对 DNA 损伤剂的敏感性或耐药性。通过挖掘该数据集，我们发现ERCC6L2 （在骨髓衰竭综合征中发生突变）编码典型的非同源末端连接途径因子，RNA 聚合酶 II 组件 ELOF1 调节对转录阻断剂的反应，并且G-四链体配体吡啶他汀的细胞毒性涉及将拓扑异构酶 II 捕获到 DNA 上。这张 DNA 损伤反应图为研究这一基本细胞系统提供了丰富的资源，并对癌症治疗中基因毒性药物的开发和使用具有重要意义。