DNAJC10 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the DNAJC10 gene in the NCI-H1299 human non-small cell lung cancer line. This loss-of-function model preserves the cellular heterogeneity of the parental line while ablating DNAJC10 expression across the population, offering a robust tool for studying gene function without clonal artifacts. The polyclonal format is ideal for investigating collective cellular responses to DNAJC10 ablation in the context of endoplasmic reticulum proteostasis.
The NCI-H1299 cell line derives from a lymph node metastasis of lung adenocarcinoma and is a widely used model for NSCLC. These epithelial cells display dysregulated proliferation and apoptosis resistance, accompanied by an adaptive reliance on endoplasmic reticulum (ER) stress pathways. Their metastatic origin and common genetic lesions make them particularly relevant for studying the intersection of oncogenic signaling and protein quality control mechanisms.
DNAJC10 (ERdj5) is an ER-luminal Hsp40 co-chaperone that catalyzes disulfide bond reduction in misfolded proteins, facilitating their retrotranslocation and proteasomal degradation via the ER-associated degradation (ERAD) pathway. Under ER stress, its expression is induced by the UPR transcription factors XBP1, ATF6, and ATF4. DNAJC10 interacts with HSPA5/BiP, EDEM1, OS9, and the HRD1/SEL1L ubiquitin ligase complex at the SEC61 translocon. These interactions target misfolded clients for degradation, alleviating ER stress and suppressing apoptotic signaling. Thus, DNAJC10 operates at a critical node linking UPR sensing to ERAD execution.
In NCI-H1299 cells, DNAJC10 knockout compromises ER quality control, potentially sensitizing the cells to ER stress-induced apoptosis. Since NSCLC tumors frequently upregulate UPR components for survival, this knockout model allows direct interrogation of DNAJC10’s role in maintaining proteostasis and promoting tumor cell fitness. Researchers can examine whether DNAJC10 loss exacerbates ER stress, alters substrate clearance, or enhances sensitivity to chemotherapeutic agents, providing insights into lung cancer vulnerabilities.
Key applications include Western blotting for UPR markers (BiP, CHOP), RT-qPCR analysis of XBP1 splicing and ATF4 targets, and flow cytometry for apoptosis (Annexin V/PI). The cells support immunofluorescence studies of ER morphology, co-immunoprecipitation of DNAJC10 with BiP, and functional assays using tunicamycin or proteasome inhibitors to assess ER stress resilience. These polyclonal knockout cells are also suitable for drug screens targeting UPR signaling in NSCLC. For further technical inquiries, contact Ascent Research.
