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• W4386252747 abstract "Abstract Interferon Stimulated Genes (ISGs) play key roles in the control of viral replication and dissemination. 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 KUNV ), 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." @default.
• W4386252747 created "2023-08-30" @default.
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• W4386252747 date "2023-08-28" @default.
• W4386252747 modified "2023-09-27" @default.
• W4386252747 title "The utility of CRISPR activation as a platform to identify interferon stimulated genes with anti-viral function" @default.
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• W4386252747 doi "https://doi.org/10.1101/2023.08.28.555046" @default.
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