bioRxiv Science⌕ Search

bioRxiv · 10.1101/2021.06.04.447090

Genome-scale CRISPR Screens Identify Host Factors that Promote Human Coronavirus Infection

Abstract

The COVID-19 pandemic has resulted in 153 million infections and 3.2 million deaths as of May 2021. While effective vaccines are being administered globally, there is still a great need for antiviral therapies as potentially antigenically distinct SARS-CoV-2 variants continue to emerge across the globe. Viruses require host factors at every step in their life cycle, representing a rich pool of candidate targets for antiviral drug design. To identify host factors that promote SARS-CoV-2 infection with potential for broad-spectrum activity across the coronavirus family, we performed genome-scale CRISPR knockout screens in two cell lines (Vero E6 and HEK293T ectopically expressing ACE2) with SARS-CoV-2 and the common cold-causing human coronavirus OC43. While we identified multiple genes and functional pathways that have been previously reported to promote human coronavirus replication, we also identified a substantial number of novel genes and pathways. Of note, host factors involved in cell cycle regulation were enriched in our screens as were several key components of the programmed mRNA decay pathway. Finally, we identified novel candidate antiviral compounds targeting a number of factors revealed by our screens. Overall, our studies substantiate and expand the growing body of literature focused on understanding key human coronavirus-host cell interactions and exploit that knowledge for rational antiviral drug development. One Sentence SummaryGenome-wide CRISPR screens identified host factors that promote human coronavirus infection, revealing novel antiviral drug targets.

Source connections

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Grodzki, M., Bluhm, A. P., Schafer, M., Tagmount, A., Russo, M., Sobh, A., Rafiee, R., Vulpe, C. D., Karst, S. M., Norris, M. H.. 2021-06-04. Genome-scale CRISPR Screens Identify Host Factors that Promote Human Coronavirus Infection. https://doi.org/10.1101/2021.06.04.447090

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Polyphyletic origins and lineage-specific expansion of chemosensory systems in phylum Actinomycetota

Phylum Actinomycetota (syn. Actinobacteriota or Actinobacteria), one of the largest well-studied bacterial phyla, is renowned for complex morphology, non-motility, ecological versatility, and secondary metabolite production. Chemosensory systems (CSS) are among the most sophisticated prokaryotic signal transduction architectures, yet their evolutionary distribution in this phylum remains poorly understood. Here, a comparative genomic analysis of 693 high-quality genomes spanning the phylum resolves the diversity, distribution, and evolutionary trajectories of CSS. Despite extensive genomic complexity and ecological versatility, only 6.9% of genomes encode complete CSS clusters, confined to a subset of lineages within a few classes, indicating lineage-specific retention or loss. In contrast, two-component systems (TCS) are ubiquitously abundant, with hybrid kinases enriched in CSS-deficient classes, suggesting functional compensation for the absence of dedicated chemosensory pathways. Basal lineages retain simplified CSS (F1 or F5), whereas Actinomycetes show lineage-specific expansion into multiple classes (F5, F8, F9, and ACF), often co-occurring, indicative of enhanced sensory complexity. The phylogeny of CheA homologs shows a polyphyletic distribution, supporting a mosaic evolutionary history shaped by multiple independent acquisitions, diversification, and gene loss. Collectively, our findings show that, unlike motile bacteria, members of Actinomycetota rely predominantly on TCS-driven regulatory networks, with CSS retained only in select lineages where ecological demands require enhanced environmental sensing. This work provides a phylum-wide evolutionary framework for the emergence, diversification, and functional specialization of CSS in non-motile bacteria.

microbiology↗

Integrating Microbial Ecology and Sustainability: Linking Agricultural Waste Gas Streams to Microbial Community Shifts in a Photobioreactor

Photobioreactors (PBRs) coupled to animal housing offer potential for mitigating gaseous emissions such as CO2 and NH3 while simultaneously producing biomass. Here, we present a time-resolved characterization of microbial community dynamics, with a focus on the genetic potential for carbon and nitrogen metabolism. Using a cone-shaped, helical PBR cultivating Arthrospira platensis (Spirulina) and supplied with exhaust air from a chicken house, we characterized microbial communities over continuous exhaust-air exposure and periodic biomass harvesting. Microbiome and gene-centric profiling revealed temporal variation in community composition and in the representation of genes associated with carbon and nitrogen metabolism. Cycle-based dissolved inorganic nitrogen balances showed that the decrease in dissolved inorganic nitrogen was only partially recovered in harvested biomass, while ammonia-oxidizing microorganisms were associated with structured, low-light niches. These observations indicate ammonia-oxidizing potential but do not establish a quantitatively relevant nitrification flux. Structured carrier matrices provided attached-growth niches harboring microbial populations underrepresented in the circulating suspension, while pre-colonized carriers facilitated biomass development during re-inoculation. Overall, our results show that coupling PBRs to agricultural exhaust streams can support biomass production while sustaining spatially structured microbial communities with taxonomic and functional potential. These findings may inform their further development for emission mitigation and circular biomass production.

microbiology↗

Engineered DNA materials shape microbial physiology and ecological selection

Extracellular DNA (eDNA) is both a structural component of microbial biofilms and a potential reservoir of genetic information, yet its role as an ecological material remains largely unexplored. Here we establish engineered extracellular materials as determinants of microbial physiology and community assembly across increasing biological complexity. A three-dimensional DNA hydrogel altered bacterial stress-associated responses, elicited species-dependent phenotypes and transcriptional changes in a defined consortium, and shaped community succession in wastewater-derived microbiomes. Distinct responses to surface-bound oligonucleotides reveal that DNA presentation matters beyond its mere presence. Strikingly, metagenomic sequencing identified rare, hydrogel-specific candidate junctions linking material-derived DNA to microbial genomic sequences, suggesting a previously unexplored interface between extracellular DNA materials and microbial genetic information. These findings reveal a dual role for engineered DNA: its material organization shapes microbial physiology and ecological selection, while its encoded sequences may engage with microbial genomic processes.

microbiology↗