The DNAJC10 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population in which the DNAJC10 gene has been disrupted. This product is built upon the NCI-H1975 human lung adenocarcinoma cell line and provides a loss-of-function model to investigate DNAJC10-dependent cellular processes. The polyclonal format maintains genetic heterogeneity while ensuring robust target gene disruption, suitable for pooled population studies without assumptions of clonal uniformity.
The NCI-H1975 host cell line is an epithelial cell model derived from a 56-year-old female nonsmoker with metastatic lung adenocarcinoma. These cells harbor the EGFR L858R/T790M double mutation, conferring resistance to first- and second-generation EGFR tyrosine kinase inhibitors (TKIs), a hallmark of acquired resistance in non-small cell lung cancer (NSCLC). NCI-H1975 is widely employed in NSCLC research, particularly for studying EGFR TKI resistance mechanisms, oncogenic signaling, and tumor cell survival under therapeutic stress.
DNAJC10 (ERdj5) is an ER co-chaperone with thioredoxin domains that reduce disulfide bonds in misfolded proteins, a prerequisite for their ER-associated degradation (ERAD). It functions within a complex containing the lectins OS9 and EDEM1, the adaptor SEL1L, and the E3 ligase HRD1 (SYVN1), and interacts with BiP (GRP78) and EDEM3. DNAJC10 is transcriptionally regulated by UPR mediators including ATF6, spliced XBP1, and IRE1??, and is induced by ER stressors like tunicamycin and thapsigargin. Its oxidoreductase activity facilitates retrotranslocation and proteasomal degradation of substrates such as mutant alpha-1 antitrypsin and misfolded MHC class I, thereby alleviating ER stress and maintaining proteostasis.
In NCI-H1975 cells, constitutive ER stress driven by oncogenic EGFR signaling and TKI resistance makes DNAJC10 likely critical for survival by enhancing clearance of misfolded proteins. Disrupting DNAJC10 may sensitize these cells to ER stress-induced apoptosis or proteasome inhibition, offering a model to dissect the interplay between protein quality control and drug resistance. This knockout population enables assessment of how loss of DNAJC10 alters UPR signaling, ERAD efficiency, and sensitivity to proteostasis perturbations, directly relevant to adaptive mechanisms in NSCLC.
Applications include western blotting for ERAD substrate accumulation, RT-qPCR for UPR gene expression analysis, flow cytometry for apoptosis under ER stress, drug sensitivity profiling with proteasome inhibitors or tunicamycin, and migration/invasion assays to assess phenotypic changes. The model supports investigation of ER stress adaptation, redox homeostasis, and chemoresistance. For further information, contact Ascent Research.
