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Genome-wide loss-of-function screen using human pluripotent stem cells to study virus-host interactions for

Ariel Pagis1, Or Alfi2, Shay Kinreich1

  • 1The Azrieli Center for Stem Cells and Genetic Research, Department of Genetics, The Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.

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|September 13, 2023
PubMed
Summary

Researchers identified novel host factors influencing SARS-CoV-2 infection using human embryonic stem cells. This study reveals MAFG and TMEM86A as key genes conferring resistance to the virus, aiding in understanding host-virus interactions.

Keywords:
Genome-wide screeningHuman pluripoptent stem cellsSARS-CoV-2

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Area of Science:

  • Virology
  • Genetics
  • Stem Cell Biology

Background:

  • Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes COVID-19, a global health crisis.
  • Understanding virus-host interactions is crucial for developing effective interventions.
  • Human embryonic stem cells (hESCs) offer a relevant model for studying host factors.

Purpose of the Study:

  • To identify host genes modulating SARS-CoV-2 infection using a genome-wide screen.
  • To discover novel host factors beyond the known ACE2 receptor.
  • To validate potential new modulators of viral entry and replication.

Main Methods:

  • Conducted a loss-of-function genome-wide screen in haploid human embryonic stem cells (hESCs).
  • Differentiated hESCs into definitive endoderm (DE) cells, which are sensitive to SARS-CoV-2.
  • Utilized CRISPR-mediated mutagenesis for gene validation and subsequent viral infection assays.

Main Results:

  • Identified the established ACE2 entry receptor as a host factor.
  • Discovered novel host factors, including transcription factor MAFG and transmembrane protein TMEM86A.
  • Validated MAFG and TMEM86A as genes conferring resistance to SARS-CoV-2 infection in differentiated DE cells.

Conclusions:

  • Genome-wide screening in hESCs is a powerful platform for identifying SARS-CoV-2 host factors.
  • MAFG and TMEM86A represent novel targets for modulating SARS-CoV-2 susceptibility.
  • This research provides insights into host-pathogen interactions in human disease-relevant cells.