Genome-wide CRISPR screening identifies new regulators of glycoprotein secretion.

Stephanie Popa, Julien Villeneuve, Sarah Stewart, Esther Perez Garcia, Anna Petrunkina Harrison, Kevin Moreau

OAI: oai:www.repository.cam.ac.uk:1810/303254 • DOI: 10.17863/CAM.50333

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.

CRISPR screen Golgi Endoplasmic Reticulum secretion