The DNAJB2 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the AGS human gastric adenocarcinoma cell line. This product provides a loss-of-function model for DNAJB2, generated through CRISPR/Cas9-mediated gene disruption, enabling the study of DNAJB2-dependent processes in a gastric cancer epithelial background. This polyclonal population retains genetic heterogeneity, reflecting a pool of edited cells, making it suitable for bulk functional assays and pooled analyses.
The AGS cell line was derived from a primary gastric adenocarcinoma resected from a 54-year-old Caucasian female and exhibits epithelial morphology. As a widely employed model of gastric adenocarcinoma, AGS cells are instrumental in dissecting signaling pathways, drug responses, and tumor biology associated with gastric cancer. The epithelial origin and tumorigenic phenotype make these cells particularly relevant for investigating chaperone-mediated proteostasis in the context of gastric malignancies.
DNAJB2 encodes a member of the Hsp40/DnaJ co-chaperone family that functions as a co-chaperone for Hsp70 (HSPA1A/HSPA8). DNAJB2 recruits Hsp70 to misfolded client proteins such as tau (MAPT) and SOD1, facilitating either refolding or, in cooperation with the E3 ubiquitin ligase STUB1/CHIP, ubiquitination and subsequent degradation by the 26S proteasome. Its activity is induced by cellular stress via the transcription factor HSF1 and neurotrophic signaling (e.g., NGF). This chaperone cycle is coupled to protein quality control pathways, including the ubiquitin-proteasome system and the unfolded protein response, with BAG3 cooperating in substrate sorting. Knockout of DNAJB2 disrupts this network, leading to accumulation of ubiquitinated proteins and impaired cellular stress resilience.
In gastric adenocarcinoma, dysregulation of proteostasis contributes to tumorigenesis and chemoresistance. DNAJB2 knockout in AGS cells provides a platform to interrogate how loss of this co-chaperone affects cancer cell viability, protein aggregation, and sensitivity to proteotoxic stress. Given the reliance of cancer cells on chaperone networks, this model is particularly suited for evaluating the impact of DNAJB2 deficiency on responses to proteasome inhibitors like bortezomib, which are investigated in gastric cancer therapy. It also offers a system to study the interplay between Hsp70-mediated protein folding and malignant transformation in an epithelial gastric context.
Researchers can employ this knockout model in a variety of downstream applications, including western blotting and RT-qPCR to assess changes in chaperone and stress-response markers, immunofluorescence to visualize ubiquitinated protein aggregates, and proteasome activity assays. Functional studies such as clonogenic survival assays and apoptosis analysis by flow cytometry facilitate examination of the knockout??s effect on cell fitness. Drug sensitivity testing with proteasome inhibitors or other chemotherapeutic agents can reveal vulnerabilities conferred by DNAJB2 loss. These applications support research into chaperone biology, proteostasis, and gastric cancer therapy. For further technical details, please contact Ascent Research.
