The DNAJC6 Knockout HT29 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population in the HT29 human colorectal adenocarcinoma cell line, with targeted disruption of the DNAJC6 gene. This loss-of-function model abolishes auxilin expression across a heterogeneous pool of edited cells, enabling pooled analysis of endocytic defects without clonal selection. The polyclonal format preserves genetic diversity, reducing the risk of clonal adaptation artifacts while maintaining a robust knockout phenotype suitable for biochemical and cell-based assays.
HT29 is an adherent epithelial cell line derived from a 44-year-old female with colorectal adenocarcinoma. It harbors well-characterized mutations in the APC tumor suppressor and the BRAF(V600E) oncogene, resulting in constitutive activation of the MAPK pathway. HT29 cells are widely employed as a model for intestinal epithelial biology, mucin production, and colorectal cancer signaling, offering a relevant context for studying how endocytic trafficking intersects with oncogenic signaling networks.
DNAJC6 encodes the co-chaperone auxilin, which functions as a key regulator of clathrin-mediated endocytosis. Auxilin specifically recruits the HSC70 (HSPA8) chaperone to clathrin-coated vesicles, where it stimulates ATP hydrolysis, thereby triggering disassembly of the clathrin lattice and releasing cargo for further trafficking. This process is essential for synaptic vesicle recycling in neurons, but also governs general endocytic events in many cell types. The auxilin/HSC70 axis interacts closely with clathrin heavy chain, dynamin (DNM1/2), the AP-2 adaptor complex, synaptojanin (SYNJ1), and endophilin (SH3GL2), coordinating vesicle scission, uncoating, and downstream receptor sorting. Disruption of auxilin function therefore impacts a core endocytic machinery, with consequences for receptor internalization, signaling attenuation, and membrane homeostasis.
In HT29 cells, knockout of DNAJC6 is predicted to impair clathrin uncoating, leading to accumulation of clathrin-coated structures and defective internalization of key receptors such as transferrin receptor and EGFR. Given that HT29 cells depend on BRAF(V600E)-driven signaling, impaired endocytic turnover of receptor tyrosine kinases could alter MAPK pathway dynamics and influence cancer cell behavior, including proliferation and drug response. This model thus provides a unique tool to dissect how endocytic defects intersect with oncogenic signaling in a colorectal cancer background, and to explore the cellular consequences of disrupted clathrin machinery.
Researchers can employ the DNAJC6 Knockout HT29 Polyclonal Cells for a broad range of applications, including transferrin uptake and EGFR internalization assays, Western blotting for clathrin and HSC70, immunofluorescence staining of clathrin puncta, live-cell imaging of endocytosis, receptor recycling assays, surface biotinylation studies, and drug sensitivity profiling. The polyclonal population is particularly suited for pooled functional genomics screens or pharmacological studies where maintaining genetic heterogeneity is advantageous. For further technical details or ordering information, please contact Ascent Research.
