The EDEM3 Knockout HGC-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population featuring disruption of the EDEM3 gene in the HGC-27 human gastric carcinoma cell line. This loss-of-function model enables investigation of endoplasmic reticulum-associated degradation (ERAD) and glycoprotein quality control without requiring single-cell cloning, offering a heterogeneous pool for robust functional assays.
HGC-27 is a widely used human gastric cancer cell line originating from the lymph node metastasis of a poorly differentiated gastric adenocarcinoma. As an epithelial cell model, it exhibits hallmarks of gastric adenocarcinoma, including aberrant growth signaling and metastatic capacity, making it valuable for investigating tumor cell biology, ER stress, and glycoprotein processing.
EDEM3 functions as an ER alpha-1,2-mannosidase that trims a terminal mannose from Man8GlcNAc2 on misfolded glycoproteins, generating a Man7GlcNAc2 signal recognized by the ERAD lectin OS9. This recognition facilitates interaction with calnexin, SEL1L, and HRD1, directing substrates to the HRD1 retrotranslocation complex for ubiquitin-proteasomal degradation. EDEM3 expression is transcriptionally activated by XBP1 (downstream of IRE1alpha) and ATF6 during ER stress, integrating its function with the unfolded protein response (UPR). By promoting clearance of misfolded glycoproteins, EDEM3 prevents toxic aggregate accumulation and maintains ER proteostasis.
In the HGC-27 background, EDEM3 knockout allows dissection of ERAD??s role in gastric cancer cell viability and metastatic behavior. Cancer cells frequently confront elevated ER stress, and loss of EDEM3 may exacerbate proteotoxic burden, impair cell growth, and enhance sensitivity to ER stress-inducing agents. This model also facilitates study of potential compensatory actions by EDEM1/EDEM2 and the broader ERAD network, including interactions with the SEL1L-HRD1 complex. Thus, it provides a physiologically relevant platform for exploring EDEM3 as a therapeutic target and for screening ER stress modulators.
Typical applications include western blotting for EDEM3 and UPR markers, RT-qPCR of XBP1/ATF6 targets, viability and apoptosis assays with tunicamycin or thapsigargin, migration/invasion studies, glycoproteomics, and cycloheximide chase analysis of glycoprotein stability. The polyclonal nature supports robust and reproducible results across experimental replicates, making it suitable for high-throughput screening of ER stress modulators. For further details or to discuss custom applications, please contact Ascent Research.
