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bioRxiv · 10.1101/2025.10.27.684930

CRISPR screen identifies CNIH1 as a selective driver of GPCR export

Assoumou, K.·Drugachenok, P.·Arrate, I. M.·Zhang, X.·Braun, S. M. G.·Stoeber, M.
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Abstract

G protein-coupled receptors (GPCRs), the largest family of transmembrane proteins, transduce extracellular stimuli into intracellular signaling cascades to orchestrate human physiology. The transport of newly synthesized receptors from the endoplasmic reticulum (ER) to the plasma membrane (PM) determines cellular responsiveness to incoming ligands, yet the molecular machinery governing GPCR export remains incompletely defined. Here, we combine a synchronized cargo-release assay with a genome-wide CRISPR/Cas9 screen to systematically map regulators of GPCR ER-to-PM transport. Focusing on the {delta}-opioid receptor (DOR), a prototypical class A GPCR, we identify CNIH1 as a dedicated export factor. In the absence of CNIH1, DOR is retained intracellularly with immature glycosylation, and drives reduced PM signaling. CNIH1 localizes to both ER exit sites and the Golgi, promoting the anterograde transport of a subset of class A GPCRs. Opioid receptors directly interact with CNIH1 and require its putative COPII-binding site for export. Distinct from other human cornichon homologs, CNIH1 defines a selective GPCR-sorting receptor that couples GPCR biosynthesis to signaling competence.

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Assoumou, K., Drugachenok, P., Arrate, I. M., Zhang, X., Braun, S. M. G., Stoeber, M.. 2025-10-28. CRISPR screen identifies CNIH1 as a selective driver of GPCR export. https://doi.org/10.1101/2025.10.27.684930

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