The DNAJC6 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population generated from the human renal clear cell adenocarcinoma line 786-O. This model features targeted disruption of the DNAJC6 gene, which encodes the clathrin-uncoating ATPase auxilin. Auxilin acts as a co-chaperone for Hsc70 (HSPA8) in clathrin-mediated endocytosis. The polyclonal nature provides a heterogeneous mix of edited cells, avoiding clonal bias and enabling robust functional analysis of DNAJC6 loss-of-function.
The 786-O cell line is derived from a human renal cell adenocarcinoma and serves as a widely used model for clear cell renal cell carcinoma (ccRCC). 786-O cells harbor a VHL mutation, leading to constitutive HIF stabilization and mimicking the hypoxic tumor microenvironment. This epithelial cancer background is valuable for studying endocytic trafficking and receptor signaling pathways that are frequently dysregulated in cancer.
DNAJC6-encoded auxilin orchestrates clathrin uncoating by recruiting Hsc70 to clathrin-coated pits. It directly interacts with clathrin heavy chain (CLTC) and the AP2 adaptor complex (AP2A1/AP2B1) to facilitate the internalization of cargo such as transferrin and epidermal growth factor receptor (EGFR). Downstream, auxilin activity is critical for EGFR degradation, transferrin recycling, and synaptic vesicle dynamics. Consequently, disruption of DNAJC6 impairs clathrin-mediated endocytosis, perturbing receptor trafficking and signaling downstream of the transferrin receptor, EGFR, and LDL receptor.
In the 786-O renal carcinoma context, DNAJC6 knockout provides a unique tool to probe the intersection of endocytosis and oncogenic signaling. Since EGFR and transferrin receptor trafficking is central to cell proliferation and iron homeostasis, auxilin loss may uncover vulnerabilities in receptor degradation pathways that are exploited by cancer cells. This model allows dissection of how disrupted endocytosis affects cancer cell viability, migration, and response to therapeutics in a VHL-mutant background.
Applications include mechanistic studies of clathrin-mediated endocytosis using transferrin uptake assays and immunofluorescence for clathrin puncta, as well as Western blotting for auxilin, Hsc70, and clathrin. The model is also suitable for Parkinson disease research, particularly PARK19-linked endocytic dysfunction, and for drug screening targeting synucleinopathies or endocytic modulators. Complementary cell viability and migration assays can assess functional outcomes. For detailed protocols and validation data, please contact Ascent Research.
