The DNAJC16 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human NCI-H1975 lung adenocarcinoma epithelial cell line, featuring targeted disruption of the DNAJC16 gene. This heterogeneous pool of cells contains various indels at the target locus, enabling loss-of-function studies of DNAJC16 in a physiologically relevant cancer background. The polyclonal format preserves population-level heterogeneity and is suitable for pooled functional screens and bulk biochemical assays.
The NCI-H1975 cell line was established from the pleural effusion of a female non-small cell lung cancer patient and harbors the EGFR T790M/L858R double mutation. It is a widely employed model for EGFR-mutant lung adenocarcinoma and drug resistance, particularly to tyrosine kinase inhibitors. Constitutive EGFR signaling drives survival, making these cells ideal for probing factors that influence mutant EGFR stability and downstream outputs.
DNAJC16 is a DnaJ domain co-chaperone that modulates Hsp70 ATPase activity to regulate protein folding, trafficking, and endoplasmic reticulum quality control. It interacts with Hsp70, Hsp90, and the ERAD component Derlin-1. Transcriptionally, DNAJC16 is regulated by HSF1 and the UPR sensors XBP1 and ATF6, linking it to proteotoxic stress. Downstream, it impacts stability of Hsp70 client proteins, such as mutant EGFR, and influences apoptosis via BCL2 family members. Thus, DNAJC16 operates at the interface of EGFR signaling and the proteostasis network.
In NCI-H1975 cells, DNAJC16 knockout is predicted to compromise Hsp70 chaperone function, causing misfolding and degradation of client proteins like oncogenic EGFR. Resulting proteostatic stress can activate the UPR (GRP78, CHOP) and alter BCL2-dependent apoptosis, potentially sensitizing cells to ER stress. This model allows dissection of how co-chaperone-mediated quality control intersects with oncogenic addiction and drug response in EGFR-mutant lung cancer.
Research applications include studying chaperone-mediated protein quality control in cancer, functional genomics screening for synthetic vulnerabilities, and exploring EGFR inhibitor resistance mechanisms. Key assays include western blotting for EGFR and apoptosis markers, RT-qPCR for UPR genes, co-immunoprecipitation of DNAJC16-Hsp70 complexes, phospho-EGFR analysis, and viability assays under ER stress. The polyclonal population is well-suited for short-term phenotypic screens. For further technical details, please contact Ascent Research.
