Key result
CRISPR activation prevents viral-induced cytopathic cell death across multiple viruses by inducing targeted ISGs.
Why the study?
Deficiencies exist in traditional plasmid-based exogenous promoter-driven expression systems used to investigate the antiviral function of interferon stimulated genes.
CRISPR activation provides a rapid and powerful methodology to identify and characterize interferon-stimulated genes and viral restriction factors from their endogenous promoters.
Plasmid-based ISG methods have deficiencies; leaves open optimized antiviral strategies pending validation.
Interferon Stimulated Gene (ISG) expression plays a key role in the control of viral replication and development of a robust adaptive response. Understanding this dynamic relationship between the pathogen and host is critical to our understanding of viral life-cycles and development of potential novel anti-viral strategies. Traditionally, plasmid based exogenous prompter driven expression of ISGs has been used to investigate anti-viral ISG function, however there are deficiencies in this approach. To overcome this, we investigated the utility of CRISPR activation (CRISPRa), which allows for targeted transcriptional activation of a gene from its endogenous promoter. Using the CRISPRa-SAM system to induce targeted expression of a panel of anti-viral ISGs we showed robust induction of mRNA and protein expression. We then employed our CRISPRa-SAM ISG panel in several antiviral screen formats to test for the ability of ISGs to prevent viral induced cytopathic cell death (CPE) and replication of Dengue Virus (DENV), Zika Virus (ZIKV), West Nile Virus Kunjin (WNV KUN ), Hepatitis A Virus (HAV) and Human Coronavirus 229E (HCoV-229E). Our CRISPRa approach confirmed the anti-viral activity of ISGs like IFI6, IFNβ and IFNλ2 that prevented viral induced CPE, which was supported by high-content immunofluorescence imaging analysis. This work highlights CRISPRa as a rapid, agile, and powerful methodology to identify and characterise ISGs and viral restriction factors.
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Kirby et al. (2024) studied Viral infection. CRISPR activation (CRISPRa-SAM system) was evaluated on Prevention of viral induced cytopathic cell death (CPE) and replication. CRISPR activation successfully induced targeted expression of anti-viral ISGs (like IFI6, IFNβ, and IFNλ2), preventing viral-induced cytopathic cell death across multiple virus types.
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