The EDEM3 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human gastric adenocarcinoma cell line AGS. This product provides a pooled loss-of-function model in which the EDEM3 gene is disrupted across a mixed population, enabling functional studies without clonal selection artifacts. As a polyclonal knockout pool, it captures the heterogeneity of CRISPR-mediated gene disruption, making it suitable for assaying population-level responses to EDEM3 deficiency.
The host AGS cell line is a widely used epithelial model of gastric adenocarcinoma. These cells lack functional p53, a common characteristic in gastric cancers, and display adherent epithelial morphology. AGS cells are known to engage endoplasmic reticulum (ER) stress pathways and represent a relevant system for investigating how protein quality control mechanisms, particularly the ER-associated degradation (ERAD) pathway, influence tumor cell behavior. Their gastric origin makes them valuable for studies of signaling and therapeutic vulnerabilities in gastric cancer.
EDEM3 encodes an alpha-1,2-mannosidase that resides in the ER lumen and serves as a key modulator of glycoprotein quality control. Under conditions of ER stress, transcription factors XBP1 and ATF6 upregulate EDEM3 expression. EDEM3 trims terminal mannose residues from N-glycans on misfolded glycoproteins, generating a specific oligosaccharide structure that is recognized by the lectins OS9 and SEL1L. This recognition targets the glycoprotein to the HRD1-DERL1 ubiquitin ligase complex for retrotranslocation into the cytosol, where VCP/p97 extracts the substrate and delivers it to the proteasome for degradation. Through this mechanism, EDEM3 functions within the core ERAD machinery to prevent the accumulation of misfolded proteins and maintain ER proteostasis.
In the context of AGS gastric adenocarcinoma cells, disruption of EDEM3 provides a powerful tool for investigating ERAD dependency and the unfolded protein response (UPR) in a cancer model with defective p53. Gastric tumors often exhibit heightened ER stress and rely on ERAD to handle proteotoxic burden. Loss of EDEM3 is expected to impair ERAD efficiency, leading to accumulation of misfolded glycoproteins and chronic UPR activation, which can influence cell survival, migration, and sensitivity to proteasome inhibitors such as bortezomib. This polyclonal knockout pool allows researchers to assess how EDEM3 loss impacts tumor-relevant phenotypes without clonal bias.
Typical applications include examining ER stress and UPR signaling by western blotting for BiP and CHOP, RT-qPCR for UPR targets, cycloheximide chase assays to monitor glycoprotein degradation, and immunofluorescence for ER morphology. Assays for apoptosis, proteasome activity, migration, and drug sensitivity with bortezomib are feasible. This model is suited for exploring EDEM3 in congenital disorders of glycosylation and gastric cancer ERAD biology. For ordering and inquiries, contact Ascent Research.
