A Genetic Map of the Response to DNA Damage in Human Cells

Cell. 2020 Jul 23;182(2):481-496.e21. doi: 10.1016/j.cell.2020.05.040. Epub 2020 Jul 9.


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.

Keywords: CRISPR; DNA damage; DNA repair; DNA-damaging agents; cancer therapeutics; functional genomics; genome stability; mechanism-of-action.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aminoquinolines / pharmacology
  • Animals
  • CRISPR-Cas Systems / genetics
  • Cell Line
  • Cytochrome-B(5) Reductase / genetics
  • Cytochrome-B(5) Reductase / metabolism
  • DNA Damage* / drug effects
  • DNA Helicases / genetics
  • DNA Helicases / metabolism
  • DNA Repair
  • DNA Topoisomerases, Type II / genetics
  • DNA Topoisomerases, Type II / metabolism
  • Gene Regulatory Networks / physiology*
  • Humans
  • Mice
  • Picolinic Acids / pharmacology
  • RNA, Guide, Kinetoplastida / metabolism
  • Tumor Suppressor Protein p53 / deficiency
  • Tumor Suppressor Protein p53 / genetics


  • Aminoquinolines
  • Picolinic Acids
  • RNA, Guide
  • Tumor Suppressor Protein p53
  • pyridostatin
  • Cytochrome-B(5) Reductase
  • DNA Helicases
  • ERCC6L2 protein, human
  • DNA Topoisomerases, Type II