CRISPR-based gene knockout screens reveal deubiquitinases involved in HIV-1 latency in two Jurkat cell models

Sci Rep. 2020 Mar 24;10(1):5350. doi: 10.1038/s41598-020-62375-3.


The major barrier to a HIV-1 cure is the persistence of latent genomes despite treatment with antiretrovirals. To investigate host factors which promote HIV-1 latency, we conducted a genome-wide functional knockout screen using CRISPR-Cas9 in a HIV-1 latency cell line model. This screen identified IWS1, POLE3, POLR1B, PSMD1, and TGM2 as potential regulators of HIV-1 latency, of which PSMD1 and TMG2 could be confirmed pharmacologically. Further investigation of PSMD1 revealed that an interacting enzyme, the deubiquitinase UCH37, was also involved in HIV-1 latency. We therefore conducted a comprehensive evaluation of the deubiquitinase family by gene knockout, identifying several deubiquitinases, UCH37, USP14, OTULIN, and USP5 as possible HIV-1 latency regulators. A specific inhibitor of USP14, IU1, reversed HIV-1 latency and displayed synergistic effects with other latency reversal agents. IU1 caused degradation of TDP-43, a negative regulator of HIV-1 transcription. Collectively, this study is the first comprehensive evaluation of deubiquitinases in HIV-1 latency and establishes that they may hold a critical role.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • CRISPR-Cas Systems*
  • DNA Polymerase III / genetics
  • DNA-Binding Proteins / genetics
  • DNA-Directed RNA Polymerases / genetics
  • Deubiquitinating Enzyme CYLD / genetics
  • Deubiquitinating Enzymes / antagonists & inhibitors
  • Deubiquitinating Enzymes / genetics*
  • Endopeptidases / genetics
  • Enzyme Inhibitors / pharmacology
  • Gene Knockout Techniques / methods*
  • HIV-1 / genetics
  • HIV-1 / physiology*
  • Host-Pathogen Interactions / genetics
  • Host-Pathogen Interactions / physiology
  • Humans
  • Jurkat Cells
  • Nucleoproteins / genetics
  • Proteasome Endopeptidase Complex / genetics
  • RNA-Binding Proteins / genetics
  • Transcription Factors / genetics
  • Ubiquitin Thiolesterase / genetics
  • Virus Latency* / drug effects


  • DNA-Binding Proteins
  • Enzyme Inhibitors
  • Iws1 protein, human
  • Nucleoproteins
  • POLE3 protein, human
  • PSMD1 protein, human
  • RNA-Binding Proteins
  • TARDBP protein, human
  • Transcription Factors
  • USP14 protein, human
  • DNA-Directed RNA Polymerases
  • DNA Polymerase III
  • Endopeptidases
  • CYLD protein, human
  • Deubiquitinating Enzyme CYLD
  • Deubiquitinating Enzymes
  • UCHL5 protein, human
  • Ubiquitin Thiolesterase
  • Proteasome Endopeptidase Complex
  • ubiquitin isopeptidase