The DNAJC16 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human gastric adenocarcinoma AGS cell line. This loss-of-function model is generated by CRISPR/Cas9-mediated disruption of the DNAJC16 gene, providing a heterogeneous pool of cells with targeted gene disruption to study DNAJC16 function in a gastric cancer context.
The AGS cell line originates from a human gastric adenocarcinoma and displays adherent epithelial morphology. It is a well-established model for investigating gastric cancer biology, including tumor cell proliferation, migration, and drug response. The AGS background provides a physiologically relevant malignant epithelial environment for examining the role of co-chaperones in cancer cell stress adaptation.
DNAJC16 encodes a J-domain containing co-chaperone that stimulates the ATPase activity of heat shock protein 70 (Hsp70), thereby facilitating substrate binding and protein folding. It operates within the Hsp70 chaperone cycle, interacting with Hsp70 (HSPA1A, HSPA8), Hsp90, and co-factors such as STUB1/CHIP and BAG family proteins. DNAJC16 is transcriptionally regulated by heat shock factor 1 (HSF1) and endoplasmic reticulum (ER) stress sensors including IRE1??, PERK, and ATF6, linking it to the unfolded protein response (UPR). Downstream, DNAJC16 influences Hsp70-mediated protein folding, clearance of protein aggregates, and autophagic degradation pathways, with key autophagy markers SQSTM1/p62 and LC3. Representative pathway components include DNAJA1-4, DNAJB1, BAG2, STUB1, PSMD1, ATG5, SQSTM1, and LC3.
In AGS gastric adenocarcinoma cells, DNAJC16 may contribute to proteotoxic stress resistance and autophagy regulation, processes critical for cancer cell survival under adverse conditions. Knockout of DNAJC16 disrupts Hsp70 chaperone function, leading to accumulation of misfolded proteins, impaired proteostasis, and altered autophagic flux. This model enables dissection of how co-chaperone-mediated protein quality control influences gastric cancer cell viability, proliferation, and stress responses. Given the role of Hsp70 in oncogenic signaling, this polyclonal knockout population is a valuable tool for exploring chaperone-targeted therapeutic strategies in gastric adenocarcinoma.
Typical applications include functional characterization of DNAJC16 in protein folding networks, screening for synthetic lethal interactions with proteasome or HSP90 inhibitors (e.g., 17-AAG), and studying autophagy and ER stress signaling using assays such as Western blotting (Hsp70, LC3, p62), RT-qPCR (XBP1 splicing, CHOP), autophagy flux measurement, colony formation, and migration assays. This knockout model supports drug sensitivity profiling and investigation of DNAJC16 as a candidate therapeutic target in gastric cancer. For further information, please contact Ascent Research.
