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J Cell Biol. 2019 Nov 4;218(11):3861-3879. doi: 10.1083/jcb.201902028. Epub 2019 Sep 5.

New factors for protein transport identified by a genome-wide CRISPRi screen in mammalian cells.

Author information

1
Department of Medical Genetics, National Institute for Health Research Cambridge Biomedical Research Centre, and Cancer Research UK Cambridge Centre, University of Cambridge, Cambridge, UK.
2
University of Cambridge Metabolic Research Laboratories, Wellcome Trust-Medical Research Council Institute of Metabolic Science, University of Cambridge, Cambridge, UK.
3
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA.
4
Department of Medical Genetics, Cambridge Institute for Medical Research, University of Cambridge, Cambridge, UK.
5
Institut de Ciencies Fotoniques, Barcelona Institute of Science and Technology, Castelldefels, Spain.
6
Centre for Genomic Regulation, Barcelona Institute of Science and Technology, Barcelona, Spain.
7
Department of Photonic Investigations, Center of Advanced Laser Technologies, National Institute for Laser, Plasma and Radiation Physics, Magurele, Romania.
8
Cell Therapy Facility, Stanford Health Care, Stanford, CA.
9
Institut National de la Sante et de la Recherche Medicale U1026, Université de Bordeaux, Bordeaux, France.
10
UK Dementia Research Institute, Cambridge, UK.
11
Department of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, CA.
12
University of Cambridge Metabolic Research Laboratories, Wellcome Trust-Medical Research Council Institute of Metabolic Science, University of Cambridge, Cambridge, UK julienvilleneuve81@gmail.com.
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Contributed equally

Abstract

Protein and membrane trafficking pathways are critical for cell and tissue homeostasis. Traditional genetic and biochemical approaches have shed light on basic principles underlying these processes. However, the list of factors required for secretory pathway function remains incomplete, and mechanisms involved in their adaptation poorly understood. Here, we present a powerful strategy based on a pooled genome-wide CRISPRi screen that allowed the identification of new factors involved in protein transport. Two newly identified factors, TTC17 and CCDC157, localized along the secretory pathway and were found to interact with resident proteins of ER-Golgi membranes. In addition, we uncovered that upon TTC17 knockdown, the polarized organization of Golgi cisternae was altered, creating glycosylation defects, and that CCDC157 is an important factor for the fusion of transport carriers to Golgi membranes. In conclusion, our work identified and characterized new actors in the mechanisms of protein transport and secretion and opens stimulating perspectives for the use of our platform in physiological and pathological contexts.

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