The DNAJC6 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human gastric adenocarcinoma cell line AGS, providing a loss-of-function model for the DNAJC6 gene. This gene encodes the co-chaperone auxilin, and the polyclonal pool consists of a heterogeneous mixture of cells carrying diverse CRISPR/Cas9-mediated disruptions of the DNAJC6 locus. The format allows researchers to study auxilin-dependent processes in a pooled population without clonal selection, making it suitable for functional genomics and drug discovery applications in a gastric cancer context.
The AGS host cell line is a widely used human gastric adenocarcinoma epithelial model initially established from a patient??s stomach tumor. AGS cells retain key characteristics of malignant gastric epithelium, including robust proliferation, adherent growth, and responsiveness to growth factors. They serve as a standard platform for investigating oncogenic signaling, tumor cell migration, and pharmacological responses. Their epithelial origin makes them particularly relevant for studying receptor-mediated endocytosis, as they express the molecular machinery necessary for clathrin-dependent internalization of cell surface receptors and other cargo.
DNAJC6 encodes auxilin, a J-domain co-chaperone that specifically recruits Hsc70/HSPA8 to clathrin-coated vesicles, where it stimulates ATP-dependent uncoating. Auxilin interacts directly with clathrin heavy chain and the AP2 adaptor complex, positioning Hsc70 at the vesicle neck to disassemble the clathrin lattice. This process is triggered by endocytic stimuli and is essential for recycling clathrin triskelia and maintaining the endocytic cycle. Through its role in uncoating, auxilin governs the trafficking and downstream signaling of numerous cargo receptors, including receptor tyrosine kinases (RTKs) such as EGFR. DNAJC6 knockout abolishes auxilin expression, impairing Hsc70 recruitment and blocking clathrin uncoating, which leads to accumulation of coated vesicles, defective endocytic recycling, and altered intracellular signaling cascades dependent on endosomal sorting.
In the AGS gastric adenocarcinoma background, disruption of auxilin function offers a unique tool to dissect the contribution of clathrin-mediated endocytosis to epithelial tumor biology. Because AGS cells rely on RTK signaling for growth and survival, the DNAJC6 knockout model enables examination of how defective clathrin uncoating alters the kinetics of receptor internalization, degradation, and downstream effector activation. This system can reveal how endocytic trafficking influences oncogenic signaling networks and may uncover vulnerabilities in gastric cancer cells that depend on efficient endocytosis for sustained proliferation and migration. The model thus bridges endocytosis research with cancer cell biology.
The DNAJC6 Knockout AGS Polyclonal Cells are suited for a variety of functional assays, including western blotting to confirm loss of auxilin protein, Sanger sequencing of the targeted genomic locus, and immunofluorescence to assess clathrin distribution. Endocytic activity can be quantified by transferrin uptake assays, while downstream signaling effects can be evaluated through EGFR degradation kinetics. Phenotypic analyses such as MTT or BrdU proliferation assays and wound healing or transwell migration assays further extend the utility of this model. Applications include investigation of clathrin-mediated endocytosis in gastric cancer, RTK trafficking and signaling, drug internalization studies, and screening for endocytosis modulators. The cells also provide a cell-based system for studying auxilin-related neurodegeneration pathways. For further details or to discuss custom projects, please contact Ascent Research.
