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  • HUMAN GENOME  (2)
Keywords
  • 1
    Keywords: SURVIVAL ; CELL ; Germany ; MICROSCOPY ; screening ; GENE ; GENES ; GENOME ; RNA ; IDENTIFICATION ; ARRAYS ; HUMAN GENOME ; MIGRATION ; PHENOTYPE ; ORGANIZATION ; INTERFERENCE ; RNA INTERFERENCE ; RESOURCE ; SCIENCE ; LIFE ; TISSUE-CULTURE CELLS
    Abstract: Despite our rapidly growing knowledge about the human genome, we do not know all of the genes required for some of the most basic functions of life. To start to fill this gap we developed a high-throughput phenotypic screening platform combining potent gene silencing by RNA interference, time-lapse microscopy and computational image processing. We carried out a genome-wide phenotypic profiling of each of the similar to 21,000 human protein-coding genes by two-day live imaging of fluorescently labelled chromosomes. Phenotypes were scored quantitatively by computational image processing, which allowed us to identify hundreds of human genes involved in diverse biological functions including cell division, migration and survival. As part of the Mitocheck consortium, this study provides an in-depth analysis of cell division phenotypes and makes the entire high-content data set available as a resource to the community
    Type of Publication: Journal article published
    PubMed ID: 20360735
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  • 2
    Keywords: CELLS ; CELL ; Germany ; human ; MICROSCOPY ; PATHWAY ; screening ; PROTEIN ; PROTEINS ; COMPLEX ; COMPLEXES ; DOMAIN ; TRANSPORT ; YEAST ; ASSAY ; MEMBRANE ; HUMAN GENOME ; GOLGI-APPARATUS ; MORPHOLOGY ; FUTURE ; ESTABLISHMENT ; REGULATOR ; REGULATORS ; DOMAINS ; ENDOPLASMIC-RETICULUM ; ER ; SUBCELLULAR-LOCALIZATION ; coiled coil ; COILED-COIL ; COPII ; MATRIX PROTEINS
    Abstract: Here we describe the establishment of microscope-based functional screening assays in intact cells that allow LIS to systematically identify new proteins involved in secretory membrane traffic, and proteins that can influence the integrity of the Golgi complex. We were able to identify 20 new proteins that affected either secretory transport, Golgi morphology, or both, when overexpressed in cells. Control experiments with human orthologs to yeast proteins with a role in membrane traffic, or already well characterized mammalian regulators of the secretory pathway, confirmed the specificity and significance of our results. Proteins localized to the Golgi complex or endoplasmic reticulum (ER) showed preferential interference in Our assays. Bioinformatic analysis of the new proteins interfering with membrane traffic and/or Golgi integrity revealed broad functional variety, but demonstrated a bias towards proteins with predicted coiled-coil domains and repeat structures. Extending our approach to a much larger set of novel proteins in the future will be an important step toward a more comprehensive understanding of the molecular basis of the secretory pathway. It will also serve as an example for similar microscope-based screens addressing different biological questions
    Type of Publication: Journal article published
    PubMed ID: 15466293
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