The DNAJC6 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population in which the DNAJC6 gene has been disrupted to ablate expression of its encoded protein, auxilin 1. This knockout product is supplied as a heterogeneous pool of HeLa cells carrying targeted gene disruptions, generated using CRISPR/Cas9 technology without single-cell clonal isolation. The resulting polyclonal population enables robust loss-of-function studies while mitigating clonal artifacts, providing a physiologically relevant model for investigating DNAJC6-dependent processes.
HeLa cells are an immortalized human cervical adenocarcinoma epithelial cell line that harbors integrated HPV-18 DNA, leading to aneuploidy and a highly proliferative phenotype. Widely used as a versatile model for cancer biology and fundamental cellular processes, HeLa cells retain key components of the endocytic and secretory machinery, making them suitable for studying membrane trafficking pathways. Their robust growth characteristics and ease of genetic manipulation facilitate efficient generation of knockout populations for functional genomics applications.
DNAJC6 encodes auxilin 1, a J-domain cochaperone that specifically recruits the chaperone Hsc70 (HSPA8) to clathrin cages, stimulating ATP hydrolysis and disassembly of clathrin coats from endocytic vesicles. This uncoating activity is essential for clathrin-mediated endocytosis and subsequent recycling of membrane components. Auxilin 1 functions at the hub of a molecular network that includes direct interactions with clathrin and the AP-2 adaptor complex, and its activity is regulated by upstream signals such as neuronal activity and calcium influx. Downstream, DNAJC6 governs the internalization of the transferrin receptor (TFRC) and epidermal growth factor receptor (EGFR), as well as the dynamics of synaptic vesicle proteins like synaptojanin. Disruption of DNAJC6 therefore impairs multiple steps in endocytic trafficking and vesicle homeostasis.
In the HeLa cell context, DNAJC6 knockout serves as a valuable model to dissect the mechanistic basis of receptor-mediated endocytosis and its links to human disease. Loss of auxilin 1 disrupts clathrin-dependent internalization pathways, leading to accumulation of clathrin-coated structures and altered surface expression of key receptors such as TFRC and EGFR. These phenotypic changes are directly relevant to the pathophysiology of Parkinson disease type 19 (PARK19) and juvenile parkinsonism, where inherited mutations in DNAJC6 cause early-onset neurodegeneration. The polyclonal HeLa knockout population thus enables detailed investigation of endocytic defects and their downstream signaling consequences in a tractable, well-characterized epithelial cell system.
This knockout cell product is ideally suited for a wide range of experimental applications, including mechanistic studies of clathrin-mediated endocytosis, Parkinson??s disease modeling, and neurodegeneration research. Researchers can employ the cells in transferrin uptake assays to quantify endocytic function, immunofluorescence microscopy to visualize clathrin puncta dynamics, and Western blotting to monitor DNAJC6, Hsc70, and clathrin. The cells also facilitate drug screening campaigns aimed at identifying enhancers of clathrin uncoating or suppressors of auxilin 1 deficiency. Electron microscopy for vesicle morphology and RNA-seq for transcriptome profiling further expand utility of the model. For further details or technical assistance, please contact Ascent Research.
