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Fluorescence-based phenotypic selection allows forward genetic screens in haploid human cells.

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.

SUBMITTER: Duncan LM

PROVIDER: S-EPMC3382162 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Fluorescence-based phenotypic selection allows forward genetic screens in haploid human cells.

Duncan Lidia M LM Timms Richard T RT Zavodszky Eszter E Cano Florencia F Dougan Gordon G Randow Felix F Lehner Paul J PJ

PloS one 20120622 6

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 in ...[more]

PMID: 22745803

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