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Updated: Jul 30, 2026
Genome-wide RNAi Screening to Identify Host Factors That Modulate Oncolytic Virus Therapy
Published on: April 3, 2018
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.
Abstract:
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019, has become a global health concern. Therefore, there is an immense need to understand the network of virus-host interactions by using human disease-relevant cells. We have thus conducted a loss-of-function genome-wide screen using haploid human embryonic stem cells (hESCs) to identify genes involved in SARS-CoV-2 infection. Although the undifferentiated hESCs are resistant to SARS-CoV-2, their differentiated definitive endoderm (DE) progenies, which express high levels of ACE2, are highly sensitive to the virus. Our genetic screening was able to identify the well-established entry receptor ACE2 as a host factor, along with additional potential novel modulators of SARS-CoV-2. Two such novel screen hits, the transcription factor MAFG and the transmembrane protein TMEM86A, were further validated as conferring resistance against SARS-CoV-2 by using CRISPR-mediated mutagenesis in hESCs, followed by differentiation of mutant lines into DE cells and infection by SARS-CoV-2. Our genome-wide genetic screening investigated SARS-CoV-2 host factors in non-cancerous human cells with endogenous ACE2 expression, providing a unique platform to identify novel modulators of SARS-CoV-2 cytopathology in human cells.
Insights
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.
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.
