The DNAJC13 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HeLa cells, featuring targeted disruption of the DNAJC13 gene. This loss-of-function model is generated using CRISPR/Cas9-mediated gene disruption, providing a genetically heterogeneous pool of knockout cells suitable for studying DNAJC13-dependent processes without clonal selection artifacts.
The HeLa cell line is a widely utilized human cervical adenocarcinoma line, HPV18-positive, with epithelial morphology. HeLa cells serve as a foundational model in cancer biology and cell signaling research, offering robust growth, ease of manipulation, and extensive characterization. Their transformed phenotype and active endocytic machinery make them an ideal host for investigating intracellular trafficking and receptor dynamics.
DNAJC13 encodes a J-domain co-chaperone that is a critical component of the retromer complex, facilitating the retrieval of cargo receptors from endosomes to the trans-Golgi network. It physically interacts with retromer subunits SNX1, SNX2, VPS35, VPS26, and VPS29, as well as clathrin and Hsc70, coordinating endosomal sorting. DNAJC13 regulates the recycling of key cargoes such as LRP6, EGFR, and the transferrin receptor, thereby influencing WNT/??-catenin and growth factor signaling. Upstream, its function is modulated by WNT ligands, EGF, and TGF-??, positioning DNAJC13 as a central node linking endocytic trafficking to signal transduction. Knockout of DNAJC13 disrupts retromer-mediated recycling, leading to impaired surface receptor homeostasis and altered downstream signaling cascades.
In HeLa cells, which express active EGFR and WNT pathways, DNAJC13 knockout provides a powerful system to dissect how endosomal sorting perturbations affect oncogenic signaling and cellular responses. Given DNAJC13??s association with Parkinson??s disease and cancer, this polyclonal knockout population enables the study of receptor trafficking defects underlying neurodegeneration and tumorigenesis. The HeLa background further allows integration with existing cancer biology data, facilitating comparative analyses of endocytic pathway dependencies.
These polyclonal knockout cells are suitable for a range of endocytic trafficking studies, retromer complex functional analyses, and investigations into WNT/EGFR crosstalk. Representative assays include Western blotting to assess protein levels, immunofluorescence for receptor localization, co-immunoprecipitation to probe retromer interactions, flow cytometry for surface receptor quantification, and migration/invasion or drug sensitivity assays to evaluate phenotypic consequences. For additional information, please contact Ascent Research.
