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

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Duncan, LM; Timms, RT; Zavodszky, E; Cano, F; Dougan, G; Randow, F; Lehner, PJDate
2012-06-22Source Title
PLoS OnePublisher
PUBLIC LIBRARY SCIENCEUniversity of Melbourne Author/s
Dougan, GordonAffiliation
Microbiology and ImmunologyMetadata
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Journal ArticleCitations
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 AccessAbstract
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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