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    Fluorescence-based phenotypic selection allows forward genetic screens in haploid human cells.
    PLoS One 2012 22;7(6):e39651. Epub 2012 Jun 22.
    Cambridge Institute for Medical Research, Addenbrooke's Hospital, Cambridge, United Kingdom.
    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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    Nature 2011 Sep 7;479(7371):131-4. Epub 2011 Sep 7.
    Wellcome Trust Centre for Stem Cell Research, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK.
    Most animals are diploid, but haploid-only and male-haploid (such as honeybee and ant) species have been described. The diploid genomes of complex organisms limit genetic approaches in biomedical model species such as mice. To overcome this problem, experimental induction of haploidy has been used in fish. Read More
    Powering mammalian genetic screens with mouse haploid embryonic stem cells.
    Mutat Res 2013 Jan-Feb;741-742:44-50. Epub 2013 Jan 28.
    Department of Animal Biotechnology, Northwest A&F University, China.
    Generating homozygous mutants in mammalian cells has been complicated by their diploid genome. If one allele of an autosomal gene was disrupted, the resulting heterozygous mutant is unlikely to display a phenotype due to the existence of the other allele. Although embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) are excellent cellular models for analyzing developmental events or disease phenotypes in vitro, a direct analysis of recessive phenotypes has been limited by their diploidy. Read More
    A reporter screen in a human haploid cell line identifies CYLD as a constitutive inhibitor of NF-κB.
    PLoS One 2013 8;8(7):e70339. Epub 2013 Jul 8.
    Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA.
    The development of forward genetic screens in human haploid cells has the potential to transform our understanding of the genetic basis of cellular processes unique to man. So far, this approach has been limited mostly to the identification of genes that mediate cell death in response to a lethal agent, likely due to the ease with which this phenotype can be observed. Here, we perform the first reporter screen in the near-haploid KBM7 cell line to identify constitutive inhibitors of NF-κB. Read More
    Genetic dissection of mammalian ERAD through comparative haploid and CRISPR forward genetic screens.
    Nat Commun 2016 Jun 10;7:11786. Epub 2016 Jun 10.
    Department of Medicine, Cambridge Institute for Medical Research, Cambridge Biomedical Campus, Hills Road, Cambridge CB2 0XY, UK.
    The application of forward genetic screens to cultured human cells represents a powerful method to study gene function. The repurposing of the bacterial CRISPR/Cas9 system provides an effective method to disrupt gene function in mammalian cells, and has been applied to genome-wide screens. Here, we compare the efficacy of genome-wide CRISPR/Cas9-mediated forward genetic screens versus gene-trap mutagenesis screens in haploid human cells, which represent the existing 'gold standard' method. Read More