The DNAJC6 Knockout Huh-7 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population engineered for targeted disruption of the DNAJC6 gene within the human Huh-7 hepatocellular carcinoma line. This polyclonal format ensures a heterogeneous pool of knockout cells, reducing clonal bias and enabling population-based assessments of auxilin function in a disease-relevant hepatic background.
The host cell line, Huh-7, was established from a well-differentiated hepatocellular carcinoma of a 57-year-old male and is widely adopted as a model for liver metabolism, viral hepatitis (notably HCV replication), and hepatocellular carcinoma research. Huh-7 cells maintain key hepatocyte features, including active endocytic and secretory pathways, making them suitable for studying intracellular trafficking events.
DNAJC6 encodes the co-chaperone auxilin, which is central to clathrin-mediated endocytosis. Auxilin recruits HSPA8 (Hsc70) to clathrin-coated vesicles, stimulating ATP-dependent uncoating and vesicle recycling. This process involves interactions with clathrin heavy chain (CLTC), the AP2 adaptor complex, and dynamin (DNM1/2). Upstream, ligand-bound receptors such as the transferrin receptor initiate coat assembly, while cellular stress signals modulate auxilin activity. Downstream, efficient clathrin uncoating is critical for synaptic vesicle recycling in neurons and endosomal sorting in other cell types. Disruption of DNAJC6 leads to impaired endocytosis and is genetically linked to juvenile-onset Parkinson disease 19 and atypical parkinsonism.
In Huh-7 cells, DNAJC6 knockout provides a unique model to investigate clathrin-mediated endocytosis in a liver-derived context, where this pathway is essential for physiological processes such as transferrin and lipoprotein uptake. The knockout allows researchers to study how auxilin deficiency affects endocytic trafficking, stress responses, and potentially oncogenic properties in a hepatocellular carcinoma background, complementing neuronal models of Parkinson’s disease.
This polyclonal knockout population is suitable for a variety of assays, including quantitative transferrin uptake assays, immunofluorescence microscopy to visualize clathrin and endosomal markers, Western blotting for partner protein analysis, and electron microscopy to assess vesicle morphology. RT-qPCR can monitor transcriptional adaptations, and cell viability assays enable drug screening in the context of impaired endocytosis. This tool supports mechanistic studies of auxilin function and therapeutic target validation for Parkinsonian disorders. For further technical information, please contact Ascent Research.
