Genetic dissection of mammalian ERAD through comparative haploid and CRISPR forward genetic screens

Richard T. Timms, Sam A. Menzies, Iva A. Tchasovnikarova, Lea C. Christensen, James C. Williamson, Robin Antrobus, Gordon Dougan, Lars Ellgaard and Paul J. Lehner ()
Additional contact information
Richard T. Timms: Cambridge Institute for Medical Research, Cambridge Biomedical Campus
Sam A. Menzies: Cambridge Institute for Medical Research, Cambridge Biomedical Campus
Iva A. Tchasovnikarova: Cambridge Institute for Medical Research, Cambridge Biomedical Campus
Lea C. Christensen: University of Copenhagen
James C. Williamson: Cambridge Institute for Medical Research, Cambridge Biomedical Campus
Robin Antrobus: Cambridge Institute for Medical Research, Cambridge Biomedical Campus
Gordon Dougan: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus
Lars Ellgaard: University of Copenhagen
Paul J. Lehner: Cambridge Institute for Medical Research, Cambridge Biomedical Campus

Nature Communications, 2016, vol. 7, issue 1, 1-10

Abstract: Abstract 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. This head-to-head comparison aimed to identify genes required for the endoplasmic reticulum-associated degradation (ERAD) of MHC class I molecules. The two approaches show high concordance (>70%), successfully identifying the majority of the known components of the canonical glycoprotein ERAD pathway. Both screens also identify a role for the uncharacterized gene TXNDC11, which we show encodes an EDEM2/3-associated disulphide reductase. Genome-wide CRISPR/Cas9-mediated screens together with haploid genetic screens provide a powerful addition to the forward genetic toolbox.

Date: 2016
References: Add references at CitEc
Citations: View citations in EconPapers (3)

Downloads: (external link)
https://www.nature.com/articles/ncomms11786 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:7:y:2016:i:1:d:10.1038_ncomms11786

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/ncomms11786

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().