Fluorescence-Based Phenotypic Selection Allows Forward Genetic Screens in Haploid Human Cells
AuthorDuncan, LM; Timms, RT; Zavodszky, E; Cano, F; Dougan, G; Randow, F; Lehner, PJ
Source TitlePLoS One
PublisherPUBLIC LIBRARY SCIENCE
University of Melbourne Author/sDougan, Gordon
AffiliationMicrobiology and Immunology
Document TypeJournal Article
CitationsDuncan, 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 StatusOpen Access
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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