The DNAJC13 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the 786-O human clear cell renal cell carcinoma (ccRCC) line. This product provides a loss-of-function model for DNAJC13, an endosomal co-chaperone critical for receptor sorting. The polyclonal format ensures a diverse pool of gene-disrupted cells, avoiding clonal selection artifacts and making it well-suited for bulk functional analyses.
The 786-O parental cell line is a well-characterized renal epithelial adenocarcinoma model harboring a truncating mutation in the VHL tumor suppressor gene, leading to constitutive hypoxia-inducible factor (HIF) stabilization and a pseudohypoxic state. Employed extensively in ccRCC research, it enables studies of tumor cell signaling, drug sensitivity, and invasion. The introduction of DNAJC13 knockout in this VHL-deficient background allows dissection of endosomal trafficking contributions to cancer cell biology.
DNAJC13 recruits HSC70 to clathrin-coated pits and early endosomes, collaborating with AP-2 and the ESCRT-0 protein Hrs to sort ubiquitinated receptors like EGFR and transferrin receptor. Downstream of EGF stimulation, it governs the balance between receptor recycling and lysosomal degradation, directly impacting MAPK and AKT pathway activation. DNAJC13 thus integrates endocytic recycling, ESCRT machinery, and clathrin-mediated endocytosis to fine-tune signaling.
In VHL-mutant 786-O cells, DNAJC13 knockout disrupts normal EGFR trafficking, potentially skewing signaling networks that support renal cancer proliferation and survival. This model is particularly relevant for studying how endosomal sorting influences sensitivity to targeted therapies, including the tyrosine kinase inhibitor sunitinib and the mTOR inhibitor everolimus. Drug resistance in ccRCC often involves altered receptor degradation pathways, making this knockout system a powerful tool to identify mechanisms and therapeutic vulnerabilities.
Typical applications include Western blotting for EGFR, phospho-AKT, and phospho-ERK; EGFR degradation kinetics; transferrin uptake assays; immunofluorescence localization of HSC70 and clathrin; co-immunoprecipitation of DNAJC13 interactors; drug sensitivity profiling; and cell migration/invasion assays. The polyclonal knockout cells are a versatile resource for investigating endocytic recycling, growth factor signaling, and drug resistance in a clinically relevant ccRCC model. For additional information, please contact Ascent Research.
