This product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the AGS human gastric adenocarcinoma cell line, in which the DNAJB11 gene has been disrupted to generate a loss-of-function model for studying endoplasmic reticulum (ER) proteostasis and the unfolded protein response (UPR). The polyclonal nature of the knockout population captures a heterogeneous mix of edited alleles, providing a robust system for functional investigations without the clonal selection artifacts inherent in monoclonal lines. The targeted disruption of DNAJB11, an ER-resident co-chaperone, is achieved through CRISPR/Cas9-mediated gene editing, yielding a versatile tool for dissecting ER stress signaling in a gastric cancer epithelial context.
The AGS cell line was originally established from a human gastric adenocarcinoma and is widely employed as an epithelial model for gastric cancer research. These adherent cells retain key characteristics of gastric tumor biology, including active signaling networks relevant to proliferation, survival, and stress responses. As a model system, AGS cells are highly transfectable and amenable to CRISPR-based genome engineering, making them an ideal host for investigating the impact of gene disruptions on cancer cell pathophysiology, particularly in the context of ER stress, which is frequently dysregulated in solid tumors.
DNAJB11 encodes an ER co-chaperone that directly interacts with the major ER chaperone BiP/GRP78 (HSPA5) to facilitate protein folding, assembly, and ER-associated degradation (ERAD). It functions within a network that includes calnexin, calreticulin, SEL1L, and the E3 ubiquitin ligase HRD1. Upstream, DNAJB11 expression is regulated by the UPR transcription factors ATF6 and XBP1, and its activity is induced by ER stress stimuli such as thapsigargin or tunicamycin. Downstream, DNAJB11 supports BiP-dependent protein quality control, and its loss perturbs the balance of UPR sensors IRE1?? and PERK, potentially leading to sustained phosphorylation of eIF2?? and upregulation of CHOP. Thus, DNAJB11 serves as a critical node linking protein folding capacity to ER stress signaling outputs.
In the gastric cancer context, DNAJB11 knockout in AGS cells disrupts ER proteostasis, creating a state of chronic ER stress that can compromise cell survival, proliferation, or trigger apoptosis??phenotypes that may reflect vulnerabilities of gastric adenocarcinoma cells to proteotoxic insults. Because cancer cells often rely on heightened ER quality control to manage increased secretory demands and oncogenic stress, this knockout model provides a platform to explore how loss of an ER co-chaperone alters tumor cell fitness. It also enables investigation of potential synthetic lethal interactions or enhanced sensitivity to ER stress?Cmodulating agents, offering insights into therapeutic strategies targeting the UPR in gastric cancer.
Typical research applications include detailed examination of the UPR and ERAD mechanisms in gastric cancer using assays such as western blotting for BiP, CHOP, and phospho-eIF2??; RT-qPCR for XBP1 mRNA splicing; and apoptosis detection via Annexin V or caspase-3/7 activation. The polyclonal knockout cells are well-suited for cell viability assays under pharmacologically induced ER stress, co-immunoprecipitation to map DNAJB11 interactomes, and migration or invasion studies to assess metastatic potential. Additionally, these cells facilitate drug discovery efforts by enabling high-throughput screening for compounds that modulate ER stress pathways. For further technical details or inquiries regarding this product, please contact Ascent Research.
