Open AccessGenome-wide CRISPR screening identifies new regulators of glycoprotein secretion
Author(s) -
Stephanie Popa,
Julien Villeneuve,
Sarah E. Stewart,
Esther Perez Garcia,
Anna Petrunkina Harrison,
Kévin Moreau
Publication year - 2020
Publication title -
wellcome open research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.298
H-Index - 21
ISSN - 2398-502X
DOI - 10.12688/wellcomeopenres.15232.2
Subject(s) - golgi apparatus , secretion , biology , gene , genome , crispr , computational biology , secretory protein , secretory pathway , glycoprotein , microbiology and biotechnology , genetics , endoplasmic reticulum , biochemistry
Background: The fundamental process of protein secretion from eukaryotic cells has been well described for many years, yet gaps in our understanding of how this process is regulated remain. Methods: With the aim of identifying novel genes involved in the secretion of glycoproteins, we used a screening pipeline consisting of a pooled genome-wide CRISPR screen, followed by secondary siRNA screening of the hits to identify and validate several novel regulators of protein secretion. Results: We present approximately 50 novel genes not previously associated with protein secretion, many of which also had an effect on the structure of the Golgi apparatus. We further studied a small selection of hits to investigate their subcellular localisation. One of these, GPR161, is a novel Golgi-resident protein that we propose maintains Golgi structure via an interaction with golgin A5. Conclusions: This study has identified new factors for protein secretion involved in Golgi homeostasis.