CRISPR activation as a platform to identify interferon stimulated genes with anti-viral function.

CRISPR activation as a platform to identify interferon stimulated genes with anti-viral function.

Publication date: Jan 23, 2024

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), 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.

Concepts Keywords
Crispra Antiviral
Death CRISPR activation
Hepatitis interferon stimulated genes
Host virus
Immunofluorescence

Semantics

Type Source Name
disease VO gene
pathway KEGG Viral replication
disease IDO pathogen
disease IDO host
drug DRUGBANK Ademetionine
disease IDO replication
disease VO Dengue virus
disease VO West Nile virus

Original Article

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