The DNAJC10 Knockout NCI-H1975 Cell Line is a CRISPR/Cas9-edited knockout cell line derived from the human lung adenocarcinoma NCI-H1975 cell line. This model enables loss-of-function studies of DNAJC10, a gene encoding an ER co-chaperone involved in protein folding and the unfolded protein response (UPR). Disruption of DNAJC10 facilitates investigation of its role in ER stress signaling, redox homeostasis, and cancer cell biology.
NCI-H1975 is a non-small cell lung cancer (NSCLC) line established from a non-smoking female adenocarcinoma patient. It carries an activating EGFR double mutation (L858R/T790M) and wild-type KRAS, making it a key model for studying EGFR-targeted therapy resistance and ER proteotoxic stress in lung cancer. These cells provide an epithelial background to evaluate DNAJC10 function in a clinically relevant oncogenic setting.
DNAJC10 encodes an ER-resident protein containing a J-domain and a thioredoxin-like domain, functioning as a co-chaperone and oxidoreductase. It interacts with HSPA5/BiP and directly regulates IRE1-alpha (ERN1) signaling, a central UPR transducer. Under ER stress, ATF4 and XBP1 transcriptionally upregulate DNAJC10, which promotes protein folding and disulfide bond formation. Knockout of DNAJC10 impairs IRE1-alpha-mediated XBP1 splicing and alters downstream signaling, affecting CHOP/DDIT3 expression and ER-associated degradation (ERAD) through interactions with PDIA3 and ERAD components. This positions DNAJC10 at the intersection of ER quality control and redox maintenance.
In NCI-H1975 cells, DNAJC10 knockout allows dissection of how ER co-chaperone deficiency impacts UPR signaling in an EGFR-mutant NSCLC background. Such cancers often display high basal ER stress and depend on adaptive UPR pathways for survival. Loss of DNAJC10 may sensitize cells to ER stress-inducing agents or modulate drug resistance, providing a platform to study the balance between adaptive and pro-apoptotic UPR branches and to identify context-specific vulnerabilities.
This knockout cell line supports diverse research applications, including validation of UPR components as therapeutic targets, mechanistic studies of ER stress-induced apoptosis, and investigation of resistance mechanisms to chemotherapeutics or EGFR inhibitors. Representative assays include western blotting for GRP78, CHOP, and phospho-eIF2alpha; RT-qPCR for XBP1 splicing; cell viability and apoptosis assays following tunicamycin or thapsigargin treatment; co-immunoprecipitation of IRE1-alpha; immunofluorescence for ER morphology; and transcriptomic analysis under ER stress. For further details, please contact Ascent Research.
