The DNAJC3 Knockout AGS Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human gastric adenocarcinoma cell line AGS, engineered for targeted DNAJC3 gene disruption. This polyclonal knockout model permits stable, loss-of-function analysis of DNAJC3 within a gastric epithelial background, circumventing the limitations of transient gene silencing. The heterogeneous pool of edited cells preserves population-level diversity, offering a robust system for functional genomic studies.
The AGS cell line serves as a widely employed model of gastric adenocarcinoma, characterized by an epithelial morphology and expression of gastric lineage markers. These cells are extensively used to investigate gastric cancer biology, including responses to therapeutic agents and microenvironmental stressors that activate the unfolded protein response (UPR). As a gastric epithelial host, AGS provides a physiologically relevant context for dissecting DNAJC3-mediated ER stress signaling pathways.
DNAJC3 encodes an ER-resident co-chaperone that functions as a negative regulator of the UPR primarily by inhibiting protein kinase R (PKR/EIF2AK2) and modulating IRE1?? (ERN1) activity. It physically interacts with HSPA5/BiP and members of the HSP40 and HSP70 chaperone families, facilitating protein folding and ER-associated degradation. Under basal conditions, DNAJC3 suppresses PKR-dependent phosphorylation of eIF2??, thereby limiting the expression of pro-apoptotic CHOP (DDIT3). ER stress sensors IRE1??, PERK, and ATF6 orchestrate UPR activation, and DNAJC3 is transcriptionally induced via the IRE1??-XBP1s branch. Thus, DNAJC3 disruption unleashes PKR activity, enhances eIF2?? phosphorylation, and potentiates CHOP-mediated apoptosis upon ER perturbation.
In the AGS gastric adenocarcinoma background, DNAJC3 knockout provides a valuable platform to explore how gastric cancer cells manage ER proteostasis and survive chronic stress conditions. By removing DNAJC3-dependent inhibition of the PKR-eIF2?? axis, this model enables direct assessment of the contribution of this signaling branch to gastric cancer cell fate decisions under ER stress. It also facilitates investigation of crosstalk between the IRE1??-XBP1s arm and the translational control machinery, providing insights into UPR-driven drug resistance mechanisms relevant to gastric cancer therapy.
This polyclonal knockout cell population is suitable for diverse experimental workflows, including western blotting for phospho-eIF2?? and CHOP, RT-qPCR for HSPA5 and DDIT3, and viability or apoptosis assays using Annexin V staining. It can be employed in PKR activation studies, XBP1 splicing analyses, and RNA sequencing to map transcriptional changes. Applications extend to drug response profiling to identify whether DNAJC3 loss sensitizes AGS cells to ER stress-inducing chemotherapeutics. For detailed product information and ordering, please contact Ascent Research.
