fluorescence-based phenotypic selection allows forward genetic screens in haploid human cells荧光技术表型选择允许前锋在单倍体人类细胞的遗传屏幕.pdfVIP

Fluorescence-Based Phenotypic Selection Allows Forward Genetic Screens in Haploid Human Cells 1. 1. 1 1 2 Lidia M. Duncan , Richard T. Timms , Eszter Zavodszky , Florencia Cano , Gordon Dougan , 3 1 Felix Randow , Paul J. Lehner * 1 Cambridge Institute for Medical Research, Addenbrooke’s Hospital, Cambridge, United Kingdom, 2 The Wellcome Trust Sanger Institute, The Wellcome Trust Genome Campus, Hinxton, United Kingdom, 3 MRC Laboratory of Molecular Biology, Division of Protein and Nucleic Acid Chemistry, Cambridge, United Kingdom 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. Citation: Duncan LM, Timms RT, Zavodszky E, Cano F, Dougan G, et al. (2012) Fluorescence-Based Phenotypic Selection Allows Forward Genetic Screens in Haploid Human Cells. PLoS ONE 7(6): e39651. doi:10.1371/journal.pone.0039651 Editor: Rachel Louise Allen, University of London, St George’s, United Kingdom Received May 21, 2012; Accepted May 29, 2012; Published June 22, 2012 Copyright: 2012 Duncan et al. This is an open-access artic