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    Fluorescence-Based Phenotypic Selection Allows Forward Genetic Screens in Haploid Human Cells

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    18
    Author
    Duncan, LM; Timms, RT; Zavodszky, E; Cano, F; Dougan, G; Randow, F; Lehner, PJ
    Date
    2012-06-22
    Source Title
    PLoS One
    Publisher
    PUBLIC LIBRARY SCIENCE
    University of Melbourne Author/s
    Dougan, Gordon
    Affiliation
    Microbiology and Immunology
    Metadata
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    Document Type
    Journal Article
    Citations
    Duncan, L. M., Timms, R. T., Zavodszky, E., Cano, F., Dougan, G., Randow, F. & Lehner, P. J. (2012). Fluorescence-Based Phenotypic Selection Allows Forward Genetic Screens in Haploid Human Cells. PLOS ONE, 7 (6), https://doi.org/10.1371/journal.pone.0039651.
    Access Status
    Open Access
    URI
    http://hdl.handle.net/11343/260026
    DOI
    10.1371/journal.pone.0039651
    Abstract
    The isolation of haploid cell lines has recently allowed the power of forward genetic screens to be applied to mammalian cells. The interest in applying this powerful genetic approach to a mammalian system is only tempered by the limited utility of these screens, if confined to lethal phenotypes. Here we expand the scope of these approaches beyond live/dead screens and show that selection for a cell surface phenotype via fluorescence-activated cell sorting can identify the key molecules in an intracellular pathway, in this case MHC class I antigen presentation. Non-lethal haploid genetic screens are widely applicable to identify genes involved in essentially any cellular pathway.

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