The DNAJC13 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the DNAJC13 gene. This loss-of-function model is derived from the NCI-H1975 human non-small cell lung cancer line, offering a genetically defined system for endosomal trafficking research. The polyclonal format maintains cellular heterogeneity for population-level analyses.
The NCI-H1975 cell line is a human lung adenocarcinoma model derived from metastatic pleural effusion of a female patient. It harbors activating EGFR mutations (L858R and T790M) that drive oncogenic signaling and alter tyrosine kinase inhibitor sensitivity. As an epithelial line, it retains tumor cell features including adherent growth and metastatic potential, making it valuable for studying drug resistance and tumor progression.
DNAJC13 functions as a co-chaperone within the retromer-mediated endosomal sorting machinery. It directly interacts with core retromer components VPS35, VPS26, and VPS29, as well as sorting nexins SNX1 and SNX2, to facilitate retrograde transport of cargo receptors such as CI-M6PR and sortilin from endosomes to the trans-Golgi network. DNAJC13 also associates with hsc70 (HSPA8) and the WASH complex, connecting it to endosomal actin dynamics. Positioned downstream of mTOR signaling and upstream of lysosomal enzyme delivery, DNAJC13 is essential for endolysosomal trafficking and receptor degradation. Its loss impairs retromer-dependent sorting, disrupting EGFR degradation and autophagic flux via effects on ATG9A trafficking and the ULK1 complex.
In the NCI-H1975 background, DNAJC13 knockout allows dissection of how endosomal trafficking intersects with oncogenic EGFR signaling. Since EGFR degradation relies partly on retromer-mediated sorting, loss of DNAJC13 may perturb receptor downregulation, fostering sustained signaling and drug resistance. Impaired autophagy in these cells could further affect stress responses dysregulated in cancer. This model thus links endosomal biology to lung cancer pathophysiology, enabling studies of trafficking errors in tumor progression.
Researchers can use these polyclonal knockout cells in diverse assays, including western blotting of retromer subunits, immunofluorescence for endosomal markers (EEA1, LAMP1), and EGFR degradation kinetics. Functional studies such as LC3?II turnover, cathepsin activity, and co?immunoprecipitation of SNX1/2 enable detailed dissection of endolysosomal and autophagy pathways. The model is also suited for drug sensitivity profiling (e.g., osimertinib) and cell migration analyses. Overall, it provides a robust platform for investigating retromer function, autophagy, and lung cancer biology. For further information, please contact Ascent Research.
