A genome-wide arrayed CRISPR screen identifies PLSCR1 as an intrinsic barrier to SARS-CoV-2 entry that recent virus variants have evolved to resist

PLoS Biol. 2024 Sep 24;22(9):e3002767. doi: 10.1371/journal.pbio.3002767. eCollection 2024 Sep.

Abstract

Interferons (IFNs) play a crucial role in the regulation and evolution of host-virus interactions. Here, we conducted a genome-wide arrayed CRISPR knockout screen in the presence and absence of IFN to identify human genes that influence Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection. We then performed an integrated analysis of genes interacting with SARS-CoV-2, drawing from a selection of 67 large-scale studies, including our own. We identified 28 genes of high relevance in both human genetic studies of Coronavirus Disease 2019 (COVID-19) patients and functional genetic screens in cell culture, with many related to the IFN pathway. Among these was the IFN-stimulated gene PLSCR1. PLSCR1 did not require IFN induction to restrict SARS-CoV-2 and did not contribute to IFN signaling. Instead, PLSCR1 specifically restricted spike-mediated SARS-CoV-2 entry. The PLSCR1-mediated restriction was alleviated by TMPRSS2 overexpression, suggesting that PLSCR1 primarily restricts the endocytic entry route. In addition, recent SARS-CoV-2 variants have adapted to circumvent the PLSCR1 barrier via currently undetermined mechanisms. Finally, we investigate the functional effects of PLSCR1 variants present in humans and discuss an association between PLSCR1 and severe COVID-19 reported recently.

MeSH terms

  • Antigens, Differentiation
  • COVID-19* / genetics
  • COVID-19* / virology
  • CRISPR-Cas Systems / genetics
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics
  • HEK293 Cells
  • Humans
  • Interferons / genetics
  • Interferons / metabolism
  • SARS-CoV-2* / genetics
  • Serine Endopeptidases / genetics
  • Serine Endopeptidases / metabolism
  • Spike Glycoprotein, Coronavirus / genetics
  • Spike Glycoprotein, Coronavirus / metabolism
  • Virus Internalization*

Substances

  • leu-13 antigen
  • TMPRSS2 protein, human
  • Spike Glycoprotein, Coronavirus
  • Serine Endopeptidases
  • Interferons
  • spike protein, SARS-CoV-2
  • Antigens, Differentiation

Supplementary concepts

  • SARS-CoV-2 variants