Genome-wide CRISPR/Cas9 Screen Identifies Host Factors Essential for Influenza Virus Replication

Cell Rep. 2018 Apr 10;23(2):596-607. doi: 10.1016/j.celrep.2018.03.045.

Abstract

The emergence of influenza A viruses (IAVs) from zoonotic reservoirs poses a great threat to human health. As seasonal vaccines are ineffective against zoonotic strains, and newly transmitted viruses can quickly acquire drug resistance, there remains a need for host-directed therapeutics against IAVs. Here, we performed a genome-scale CRISPR/Cas9 knockout screen in human lung epithelial cells with a human isolate of an avian H5N1 strain. Several genes involved in sialic acid biosynthesis and related glycosylation pathways were highly enriched post-H5N1 selection, including SLC35A1, a sialic acid transporter essential for IAV receptor expression and thus viral entry. Importantly, we have identified capicua (CIC) as a negative regulator of cell-intrinsic immunity, as loss of CIC resulted in heightened antiviral responses and restricted replication of multiple viruses. Therefore, our study demonstrates that the CRISPR/Cas9 system can be utilized for the discovery of host factors critical for the replication of intracellular pathogens.

Keywords: CIC; CRISPR/Cas9 screen; Capicua; GeCKO; H5N1; SLC35A1; cell-intrinsic immunity; host factors; influenza virus; sialic acid pathway.

MeSH terms

  • A549 Cells
  • CRISPR-Cas Systems / genetics*
  • Gene Knockout Techniques / methods*
  • Gene Library
  • Genome, Human
  • Humans
  • Influenza A Virus, H5N1 Subtype / genetics
  • Influenza A Virus, H5N1 Subtype / physiology*
  • Lentivirus / genetics
  • Nucleotide Transport Proteins / genetics
  • Nucleotide Transport Proteins / metabolism
  • Virus Internalization
  • Virus Replication

Substances

  • Nucleotide Transport Proteins
  • SLC35A1 protein, human