The DNAJC6 Knockout HGC-27 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal population of the HGC-27 cell line carrying targeted disruption of the DNAJC6 gene. As a polyclonal knockout pool, it comprises a heterogeneous mixture of genotypes arising from non-clonal editing, providing a loss-of-function model that avoids the biases of single-cell-derived clones. This versatile tool allows researchers to investigate auxilin-dependent processes with population-level resolution, making it well-suited for functional genomic screens or studies where clonal variation would confound interpretation.
The parental HGC-27 cell line originates from a poorly differentiated gastric adenocarcinoma metastatic to a lymph node, representing an established in vitro system for gastric carcinoma research. These epithelial cells retain features of gastric mucosal physiology, including secretory function and barrier integrity, while displaying transformative characteristics such as altered adhesion and enhanced proliferative capacity. HGC-27 cells are extensively employed to dissect signaling pathways that converge on endocytosis, cell polarity, and tumor metastasis.
The DNAJC6 gene encodes auxilin, a J-domain co-chaperone that recruits HSPA8 (Hsc70) to clathrin-coated vesicles to drive ATP-dependent clathrin uncoating, an obligatory step in receptor-mediated endocytosis. Auxilin function is modulated by the LRRK2 kinase and involves physical interactions with the AP-2 adaptor complex and clathrin heavy chain. Consequent to DNAJC6 disruption, HSPA8 fails to efficiently localize to nascent vesicles, thus hindering clathrin coat disassembly, impairing internalization of cargo receptors, and dysregulating downstream effectors including dynamin-1 (DNM1) and synaptojanin-1 (SYNJ1).
In HGC-27 gastric carcinoma cells, DNAJC6 knockout allows dissection of how clathrin-mediated endocytosis regulates malignant traits, including proliferation, migration, and epithelial polarity. Because these cells depend on balanced receptor trafficking for growth signaling, auxilin loss may reveal targetable vulnerabilities. Additionally, this system enables mechanistic studies of LRRK2?Cauxilin signaling outside the nervous system, providing a complementary model for investigating endocytic defects linked to Parkinson disease.
Researchers can apply this polyclonal knockout model to transferrin uptake assays for bulk endocytosis, co-immunoprecipitation to verify loss of auxilin?CHSPA8 interaction, and immunofluorescence to detect clathrin-coated pit accumulation. The pool is suitable for RT-qPCR analysis of endocytic gene networks, Western blotting of pathway markers, and LRRK2 kinase activity assays under pharmacological inhibition. Therefore, this product is a versatile platform for screening endocytosis modulators or LRRK2-targeting compounds in oncology and neurodegeneration research. For further details, please contact Ascent Research.
