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Identification of Required Host Factors for SARS-CoV-2 Infection in Human Cells
Zharko Daniloski1, Tristan X Jordan2, Hans-Hermann Wessels1
1New York Genome Center, New York, NY, USA; Department of Biology, New York University, New York, NY, USA.
Abstract:
To better understand host-virus genetic dependencies and find potential therapeutic targets for COVID-19, we performed a genome-scale CRISPR loss-of-function screen to identify host factors required for SARS-CoV-2 viral infection of human alveolar epithelial cells. Top-ranked genes cluster into distinct pathways, including the vacuolar ATPase proton pump, Retromer, and Commander complexes. We validate these gene targets using several orthogonal methods such as CRISPR knockout, RNA interference knockdown, and small-molecule inhibitors. Using single-cell RNA-sequencing, we identify shared transcriptional changes in cholesterol biosynthesis upon loss of top-ranked genes. In addition, given the key role of the ACE2 receptor in the early stages of viral entry, we show that loss of RAB7A reduces viral entry by sequestering the ACE2 receptor inside cells. Overall, this work provides a genome-scale, quantitative resource of the impact of the loss of each host gene on fitness/response to viral infection.
Insights
This study identified essential host genes for SARS-CoV-2 infection using CRISPR screening. Discovering these host factors reveals new therapeutic targets for COVID-19 treatment.
Area of Science:
- Virology
- Genetics
- Cell Biology
Background:
- Understanding host-virus interactions is crucial for developing effective COVID-19 therapies.
- Identifying host genetic dependencies can reveal novel therapeutic targets.
Purpose of the Study:
- To perform a genome-scale CRISPR screen to identify host factors essential for SARS-CoV-2 infection.
- To uncover potential therapeutic targets for COVID-19 by analyzing host-virus genetic dependencies.
Main Methods:
- Genome-scale CRISPR loss-of-function screen in human alveolar epithelial cells.
- Validation using CRISPR knockout, RNA interference, and small-molecule inhibitors.
- Single-cell RNA-sequencing to analyze transcriptional changes.
Main Results:
- Identified key host pathways, including vacuolar ATPase proton pump, Retromer, and Commander complexes, required for SARS-CoV-2 infection.
- Observed shared transcriptional changes in cholesterol biosynthesis upon loss of top-ranked genes.
- Demonstrated that RAB7A depletion reduces viral entry by sequestering the ACE2 receptor.
Conclusions:
- This study provides a comprehensive resource of host genes impacting SARS-CoV-2 infection.
- The identified host factors represent potential therapeutic targets for COVID-19.
- Understanding host genetic dependencies offers insights into viral replication mechanisms.
