Genome-wide CRISPR/Cas9 Screen Identifies Host Factors Essential for Influenza Virus Replication. 2018

Julianna Han, and Jasmine T Perez, and Cindy Chen, and Yan Li, and Asiel Benitez, and Matheswaran Kandasamy, and Yoontae Lee, and Jorge Andrade, and Benjamin tenOever, and Balaji Manicassamy
Department of Microbiology, The University of Chicago, Chicago, IL 60637, USA.

The emergence of influenza A viruses (IAVs) 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 IAVs. 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.

UI MeSH Term Description Entries
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000072283 A549 Cells An immortalized cell line derived from human ADENOCARCINOMA, ALVEOLAR basal epithelial cells isolated from the lungs of a male patient in 1972. The cell line is positive for KERATIN, can synthesize LECITHIN, and contains high levels of POLYUNSATURATED FATTY ACIDS in its PLASMA MEMBRANE. It is used as a model for PULMONARY ALVEOLI function and virus infections, as a TRANSFECTION host, and for PRECLINICAL DRUG EVALUATION. A549 Cell Line,A549 Cell,A549 Cell Lines,Cell Line, A549,Cell Lines, A549,Cell, A549,Cells, A549
D014779 Virus Replication The process of intracellular viral multiplication, consisting of the synthesis of PROTEINS; NUCLEIC ACIDS; and sometimes LIPIDS, and their assembly into a new infectious particle. Viral Replication,Replication, Viral,Replication, Virus,Replications, Viral,Replications, Virus,Viral Replications,Virus Replications
D015723 Gene Library A large collection of DNA fragments cloned (CLONING, MOLECULAR) from a given organism, tissue, organ, or cell type. It may contain complete genomic sequences (GENOMIC LIBRARY) or complementary DNA sequences, the latter being formed from messenger RNA and lacking intron sequences. DNA Library,cDNA Library,DNA Libraries,Gene Libraries,Libraries, DNA,Libraries, Gene,Libraries, cDNA,Library, DNA,Library, Gene,Library, cDNA,cDNA Libraries
D015894 Genome, Human The complete genetic complement contained in the DNA of a set of CHROMOSOMES in a HUMAN. The length of the human genome is about 3 billion base pairs. Human Genome,Genomes, Human,Human Genomes
D016086 Lentivirus A genus of the family RETROVIRIDAE consisting of non-oncogenic retroviruses that produce multi-organ diseases characterized by long incubation periods and persistent infection. Lentiviruses are unique in that they contain open reading frames (ORFs) between the pol and env genes and in the 3' env region. Five serogroups are recognized, reflecting the mammalian hosts with which they are associated. HIV-1 is the type species. Lentivirinae,Lentiviruses
D053124 Influenza A Virus, H5N1 Subtype A subtype of INFLUENZA A VIRUS comprised of the surface proteins hemagglutinin 5 and neuraminidase 1. The H5N1 subtype, frequently referred to as the bird flu virus, is endemic in wild birds and very contagious among both domestic (POULTRY) and wild birds. It does not usually infect humans, but some cases have been reported. H5N1 Avian Influenza Virus,H5N1 Bird Flu Virus,H5N1 Virus,H5N1 subtype,H5N1 Viruses,H5N1 subtypes,Virus, H5N1
D053586 Virus Internalization The entering of cells by viruses following VIRUS ATTACHMENT. This is achieved by ENDOCYTOSIS, by translocation of the whole virus across the cell membrane, by direct MEMBRANE FUSION of the viral membrane with the CELL MEMBRANE, or by fusion of the membrane of infected cells with the membrane of non-infected cells causing SYNCYTIA to be formed. Viral Entry,Viral Internalization,Viral Membrane Fusion,Virus Entry,Virus Membrane Fusion,Entry, Viral,Entry, Virus,Fusion, Viral Membrane,Internalization, Viral,Internalization, Virus,Membrane Fusion, Viral
D055786 Gene Knockout Techniques Techniques to alter a gene sequence that result in an inactivated gene, or one in which the expression can be inactivated at a chosen time during development to study the loss of function of a gene. Gene Knock-Out Techniques,Gene Knock Out,Gene Knock Out Techniques,Gene Knockout,Gene Knock Outs,Gene Knock-Out Technique,Gene Knockout Technique,Gene Knockouts,Knock Out, Gene,Knock Outs, Gene,Knock-Out Technique, Gene,Knock-Out Techniques, Gene,Knockout Technique, Gene,Knockout Techniques, Gene,Knockout, Gene,Knockouts, Gene,Out, Gene Knock,Outs, Gene Knock,Technique, Gene Knock-Out,Technique, Gene Knockout,Techniques, Gene Knock-Out,Techniques, Gene Knockout
D033704 Nucleotide Transport Proteins Proteins involved in the transport of NUCLEOTIDES across cellular membranes. Nucleotide Translocase,Transport Proteins, Nucleotide

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