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A genome-wide CRISPR activation screen identifies B4GALNT2 as an inhibitory host factor for influenza virus infection

ABSTRACT: A genome-wide CRISPR activation screen identifies B4GALNT2 as an inhibitory host factor for influenza virus infection

PROVIDER: PRJNA351924 | ENA |

REPOSITORIES: ENA

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Genome-Wide CRISPR/Cas9 Screen Identifies Host Factors Essential for Influenza Virus Replication

Project description:The emergence of influenza A viruses (IAV) from zoonotic reservoirs poses a great threat to human health. As seasonal vaccines are ineffective against zoonotic strains, and newly transmitted viruses can quickly acquire drug resistance, there remains a need for host- directed therapeutics against IAV. Here, we performed a genome-scale CRISPR/Cas9 knockout screen in human lung epithelial cells with a human isolate of an avian H5N1 strain. Several genes involved in sialic acid biosynthesis and related glycosylation pathways were highly enriched post-H5N1 selection, including SLC35A1, a sialic acid transporter essential for IAV receptor expression and thus viral entry. Importantly, we have identified capicua (CIC) as a negative regulator of cell intrinsic immunity, as loss of CIC resulted in heightened antiviral responses and restricted replication of multiple viruses. Therefore, our study demonstrates that the CRISPR/Cas9 system can be utilized for the discovery of host factors critical for the replication of intracellular pathogens.

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A genome-wide CRISPR activation screen identifies B4GALNT2 as an inhibitory host factor for influenza virus infection

Project description:Identification of factors to inhibit influenza virus infection

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Genome-Wide CRISPR/Cas9 Screen Identifies Host Factors Essential for Influenza Virus Replication

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Genome-wide CRISPR screen Identifies Host Dependency Factors for Influenza A Virus Infection

Project description:In this study, we performed a genome-wide CRISPR screen in A549 cells to identify host genes that are required forInfluenza A virus (IAV) infection.Amongst the identified genes, WDR7, CCDC115, TMEM199 and CMTR1 conferred strong protection against IAV infection when they were CRISPR-deleted without impacting host cell fitness. To investigate the roles of these genes on the transciptomic states of the cell, we performed bulk RNA-sequencing of WDR7, CCDC115, TMEM199 and CMTR1 polyclonal knockout cells. We found that cells lacking WDR7, CCDC115 or TMEM199 up-regulates expression of lysosomal genes due to increased nuclear translocation of Transcription factor EB (TFEB), which hampers IAV entry. In addition,we report that CRISPR deletion of CMTR1 results in increased expression of Interferon-stimulated genes (ISGs) that also plays a role inblocking IAV infection.

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Genome-scale CRISPR screen reveals inhibitory factors as regulatory module of HIV

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A Virus-Packageable CRISPR Screen Identifies Host Factors Mediating Interferon Inhibition of HIV

Project description:Interferon (IFN) inhibits HIV replication by inducing an array of antiviral effectors. Here we describe a novel CRISPR knockout screening approach to identify the ensemble of these HIV restriction factors. We assembled a CRISPR sgRNA library specific for Interferon Stimulated Genes (ISGs) into a modified lentiviral vector that allows for packaging of sgRNA-encoding genomes in trans into budding HIV-1 particles. We observed that knockout of Zinc Antiviral Protein (ZAP) improved the performance of the screen due to ZAP-mediated inhibition of the vector. We identify a small panel of IFN-induced HIV restriction factors, including MxB, IFITM1, Tetherin/BST2 and TRIM5 which together explain the inhibitory effects of IFN on the HIV-1 LAI strain in THP-1 cells. Further, we identify novel HIV dependency factors, including SEC62 and TLR2. The ability of IFN-induced restriction factors to inhibit an HIV strain to replicate in human cells suggests that these human restriction factors are incompletely antagonized.

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