Parallel shRNA and CRISPR-Cas9 screens enable antiviral drug target identification

Details

• Alternative Title:
  Nat Chem Biol
• Personal Author:
  Deans, Richard M. ; Morgens, David W. ; Ökesli, Ayşe ; Pillay, Sirika ; Horlbeck, Max A. ; Kampmann, Martin ; Gilbert, Luke A. ; Li, Amy ; Mateo, Roberto ; Smith, Mark ; Glenn, Jeffrey S. ; Carette, Jan E. ; Khosla, Chaitan ; Bassik, Michael C.
• Description:
  Broad-spectrum antiviral drugs targeting host processes could potentially treat a wide range of viruses while reducing the likelihood of emergent resistance. Despite great promise as therapeutics, such drugs remain largely elusive. Here we used parallel genome-wide high-coverage short hairpin RNA (shRNA) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 screens to identify the cellular target and mechanism of action of GSK983, a potent broad-spectrum antiviral with unexplained cytotoxicity. We found that GSK983 blocked cell proliferation and dengue virus replication by inhibiting the pyrimidine biosynthesis enzyme dihydroorotate dehydrogenase (DHODH). Guided by mechanistic insights from both genomic screens, we found that exogenous deoxycytidine markedly reduced GSK983 cytotoxicity but not antiviral activity, providing an attractive new approach to improve the therapeutic window of DHODH inhibitors against RNA viruses. Our results highlight the distinct advantages and limitations of each screening method for identifying drug targets, and demonstrate the utility of parallel knockdown and knockout screens for comprehensive probing of drug activity.
• Subjects:
  Antiviral Agents Article Carbazoles Cell Line, Tumor Cloning, Molecular CRISPR-Cas Systems Humans Lentivirus RNA, Small Interfering
• Source:
  Nat Chem Biol. 12(5):361-366
• Pubmed ID:
  27018887
• Pubmed Central ID:
  PMC4836973
• Document Type:
  Journal Article
• Funding:
  K99 CA181494/CA/NCI NIH HHS/United States ; DP2 HD084069/HD/NICHD NIH HHS/United States ; R00 CA181494/CA/NCI NIH HHS/United States ; DP2 AI104557/AI/NIAID NIH HHS/United States ; T32 GM008284/GM/NIGMS NIH HHS/United States ; T32 GM007618/GM/NIGMS NIH HHS/United States ; K99/R00 CA181494/CA/NCI NIH HHS/United States ; 1DP2HD084069-01/DP/NCCDPHP CDC HHS/United States ; T32 EB009383/EB/NIBIB NIH HHS/United States ; U19 AI109662/AI/NIAID NIH HHS/United States ; U19-AI109662/AI/NIAID NIH HHS/United States
• Volume:
  12
• Issue:
  5
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:6595c5bbbc2167ea3e2806c12a40b628d8cabbb819e1069a4621a23cdbecc30c
• Download URL:
  https://stacks.cdc.gov/view/cdc/40994/cdc_40994_DS1.pdf
• File Type:
  [PDF - 1.17 MB ]