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      Haploid genetic screens in human cells identify host factors used by pathogens.

      Science (New York, N.Y.)

      ADP Ribose Transferases, metabolism, toxicity, Adenosine Diphosphate Ribose, Amino Acid Sequence, Antigens, Bacterial, Bacterial Toxins, Biosynthetic Pathways, Cell Line, Tumor, Diphtheria Toxin, Exotoxins, Genes, Genetic Testing, Haploidy, Histidine, analogs & derivatives, biosynthesis, Host-Pathogen Interactions, Humans, Influenza A Virus, H1N1 Subtype, pathogenicity, Molecular Sequence Data, Monosaccharide Transport Proteins, genetics, Mutagenesis, Insertional, N-Acylneuraminate Cytidylyltransferase, Peptide Elongation Factor 2, Proteins, chemistry, Virulence Factors

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          Abstract

          Loss-of-function genetic screens in model organisms have elucidated numerous biological processes, but the diploid genome of mammalian cells has precluded large-scale gene disruption. We used insertional mutagenesis to develop a screening method to generate null alleles in a human cell line haploid for all chromosomes except chromosome 8. Using this approach, we identified host factors essential for infection with influenza and genes encoding important elements of the biosynthetic pathway of diphthamide, which are required for the cytotoxic effects of diphtheria toxin and exotoxin A. We also identified genes needed for the action of cytolethal distending toxin, including a cell-surface protein that interacts with the toxin. This approach has both conceptual and practical parallels with genetic approaches in haploid yeast.

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          Journal
          19965467
          10.1126/science.1178955

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