DNAJB5 Knockout AGS Polyclonal Cells are a human polyclonal knockout cell population engineered through CRISPR/Cas9-mediated disruption of the DNAJB5 gene in AGS gastric adenocarcinoma cells. This product comprises a heterogeneous pool of edited cells, avoiding clonal selection, and thus enables population-level interrogation of DNAJB5-dependent mechanisms in a gastric cancer background. The knockout model is suited for functional proteomics, stress biology, and chaperone network studies where loss-of-function effects are assessed using bulk cell assays.
The host AGS cell line originates from a human gastric adenocarcinoma and displays an epithelial, adherent morphology. It is a widely utilized in vitro model for gastric cancer research, offering a relevant context to examine oncogenic signaling, therapeutic responses, and cellular stress pathways. The AGS background provides a clinically pertinent environment for studying how co-chaperone disruption impacts malignant cell physiology and stress adaptation.
DNAJB5 is a DNAJ/Hsp40 family co-chaperone that stimulates the ATPase activity of Hsp70 (notably HSPA1A and HSPA8), driving client protein folding and preventing aggregation. Within the chaperone network, DNAJB5 interacts with BAG family co-chaperones, the E3 ubiquitin ligase STUB1/CHIP, and the Hsp70-organizing protein HOP. Its expression is transcriptionally regulated by heat shock factor 1 (HSF1) in response to cellular stress, such as heat or oxidative challenges. DNAJB5?CHsp70 complexes recognize and process misfolded proteins, including kinases and transcription factors; therefore, DNAJB5 depletion is expected to impair Hsp70 activity, leading to accumulation of misfolded conformers and proteotoxic stress.
The DNAJB5 knockout in AGS cells likely disrupts proteostasis networks critical for gastric cancer cell survival under adverse conditions like hypoxia or nutrient deprivation. Impaired Hsp70-mediated folding may destabilize oncogenic clients, sensitizing cells to stress-induced apoptosis or altering signaling pathways that govern proliferation and drug resistance. This model facilitates dissection of how co-chaperone dysfunction influences gastric cancer fitness, offering insights into the therapeutic potential of targeting the Hsp70 cycle in gastric malignancies.
Researchers can utilize this polyclonal knockout population to study proteostasis using western blotting for Hsp70 and client proteins, co-immunoprecipitation to confirm disrupted DNAJB5?CHsp70 interactions, and thermal shift assays to monitor protein aggregation. Functional assessments may include cell viability (MTT/CCK-8) and apoptosis (Annexin V/PI) assays to evaluate stress sensitivity, alongside proteasome activity assays to explore compensatory degradation. The model also supports neurodegenerative disease research where proteostasis collapse is central. For additional information, please contact Ascent Research.
