The DNAJC6 Knockout K-562 Polyclonal Cells are a polyclonal knockout cell population generated by CRISPR/Cas9-mediated disruption of the DNAJC6 gene in the human K-562 cell line. This product provides a genetically heterogeneous pool of cells carrying targeted gene disruptions, enabling loss-of-function studies without clonal selection. The knockout model allows researchers to investigate the functional consequences of DNAJC6 ablation in a cellular context amenable to high-throughput assays and suspension culture. It is designed for applications requiring robust endocytosis models free from the complexities of neuronal systems.
K-562 cells are a widely characterized human chronic myelogenous leukemia (CML) cell line originally established from the pleural effusion of a patient in blast crisis. These cells are undifferentiated, multipotent progenitors capable of differentiating along erythroid, granulocytic, and monocytic lineages under appropriate stimuli. As a suspension cell line, K-562 offers high reproducibility and scalability for biochemical experiments, flow cytometry, and live-cell imaging. Its well-documented genetic background and robust growth make it an ideal host for studying clathrin-mediated endocytosis and receptor trafficking in a hematopoietic context.
DNAJC6 encodes auxilin, a J-domain co-chaperone that specifically recruits heat shock cognate 70 (Hsc70/HSPA8) to clathrin-coated vesicles. By stimulating Hsc70??s ATPase activity, auxilin drives the dissociation of the clathrin lattice, a critical step in vesicle uncoating and cargo internalization. Auxilin functions downstream of neuronal activity and is regulated by transcription factors such as SP1 and CREB. It interacts directly with the clathrin heavy chain (CLTC), the AP2 adaptor complex (including AP2A1 and AP2M1), and the GTPase dynamin (DNM1). Disruption of DNAJC6 impairs clathrin-mediated endocytosis and synaptic vesicle recycling, linking auxilin dysfunction to neurodegenerative disorders like Parkinson??s disease.
In the K-562 cell model, DNAJC6 knockout disrupts the normal endocytic machinery, offering a simplified system to dissect clathrin-dependent internalization pathways without neuronal synaptic specialization. Although K-562 cells lack synaptic vesicles, they express all necessary components for clathrin-mediated endocytosis, including CLTC, AP2A1, HSPA8, and DNM1. This knockout model thus enables the study of auxilin??s role in general endocytic processes, such as transferrin receptor uptake, growth factor signaling, and immune receptor internalization. Additionally, it serves as a valuable platform for investigating how impaired endocytosis may contribute to leukemia cell biology or for comparative analyses with Parkinson??s disease-related neurodegeneration.
Researchers can employ this polyclonal knockout population in a variety of assays, including western blotting and RT-qPCR to confirm gene disruption, immunofluorescence to visualize clathrin coat accumulation, and transferrin uptake assays to quantify endocytosis defects. Co-immunoprecipitation experiments can assess altered interactions between auxilin and its partners, while flow cytometry can monitor surface receptor dynamics. This model is particularly suited for drug discovery screens targeting endocytosis modulators, high-content imaging of vesicle trafficking, and functional complementation studies. For further technical details or to discuss customization options, please contact Ascent Research.
