CRISPR/CAS9-based DNA damage response screens reveal gene-drug interactions

DNA Repair (Amst). 2020 Mar:87:102803. doi: 10.1016/j.dnarep.2020.102803. Epub 2020 Jan 16.

Abstract

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.

Keywords: CRISPR screen; DNA damage response; TP53.

Publication types

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

MeSH terms

  • CRISPR-Associated Protein 9*
  • CRISPR-Cas Systems*
  • DNA Damage / genetics*
  • DNA Polymerase III / genetics
  • DNA-Binding Proteins / genetics
  • Drug Resistance / genetics*
  • Gene Library
  • Genes, p53 / genetics
  • HEK293 Cells
  • Humans
  • Nucleoproteins / genetics
  • Poly(ADP-ribose) Polymerase Inhibitors / metabolism
  • Small Molecule Libraries / pharmacology*

Substances

  • DNA-Binding Proteins
  • Nucleoproteins
  • POLE3 protein, human
  • Poly(ADP-ribose) Polymerase Inhibitors
  • Small Molecule Libraries
  • DNA Polymerase III
  • CRISPR-Associated Protein 9