The DNAJC6 Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population that provides a loss-of-function model for studying the human DNAJC6 gene. This product consists of a heterogeneous population of SK-HEP-1 cells with targeted disruption of DNAJC6, enabling robust investigation of auxilin-dependent endocytic processes in a tumorigenic cellular context. The polyclonal format maintains the diversity of editing outcomes while eliminating reliance on single-cell clones, offering a practical and reproducible tool for biochemical and cell-based assays without clonal selection artifacts.
The host cell line, SK-HEP-1, is a human hepatic adenocarcinoma cell line originally derived from the ascites of a patient with liver adenocarcinoma. These adherent epithelial-like cells exhibit tumorigenic properties and are widely employed in cancer biology and endocytosis research. SK-HEP-1 cells retain functional clathrin-mediated endocytosis machinery and express key components of the endocytic pathway, making them an ideal platform for dissecting the molecular mechanisms of endocytosis and its dysregulation in disease states.
DNAJC6 encodes auxilin, a J-domain protein that coordinates heat-shock protein 70 (Hsc70/HSPA8) recruitment to clathrin-coated vesicles. Auxilin binds directly to assembled clathrin triskelia via its clathrin-binding domain and stimulates the ATPase activity of Hsc70, driving vesicle uncoating. This process is essential for clathrin-mediated endocytosis and synaptic vesicle recycling. Key interacting factors include clathrin heavy chain (CLTC), adaptor protein complex 2 subunit alpha 1 (AP2A1), synaptojanin 1 (SYNJ1), and cyclin G-associated kinase (GAK). Auxilin functions downstream of endocytic cargo and synaptic activity and acts upstream of clathrin disassembly and vesicle recycling; loss of auxilin disrupts the uncoating cycle, leading to accumulation of clathrin-coated intermediates and impaired endocytic trafficking.
In the SK-HEP-1 host cell context, knockout of DNAJC6 disrupts clathrin-mediated endocytosis without the confounding influence of neuron-specific factors, allowing researchers to isolate core endocytic defects. The absence of functional auxilin in this hepatic adenocarcinoma line leads to defective uptake of transferrin and other cargo, which can be quantified by established assays. Although auxilin pathology is canonically associated with Parkinson??s disease and juvenile parkinsonism, the fundamental role of clathrin dynamics in cellular homeostasis makes this model broadly applicable to endocytosis mechanism studies and drug screening campaigns aimed at modulating auxilin activity or restoring vesicle trafficking. This system is particularly valuable for investigating how endocytic dysfunction contributes to neurodegeneration and for evaluating therapeutic candidates in a human cell background.
This polyclonal knockout cell population is suitable for a range of research applications including Parkinson’s disease research, endocytosis mechanism studies, and drug screening for synucleinopathies. Researchers can validate knockout by Western blotting for DNAJC6 and assess functional consequences via transferrin uptake assays, immunofluorescence staining for clathrin puncta, and RT-qPCR analysis of endocytic gene expression. Additional assays such as apoptosis and cell viability measurements facilitate the study of downstream cellular responses to endocytic impairment. For further information, please contact Ascent Research.
