The DNAJB14 Knockout NCI-H1299 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human non-small cell lung carcinoma line NCI-H1299. This product provides a loss-of-function model for DNAJB14, enabling investigation of its role as a J-domain co-chaperone. The polyclonal format consists of a heterogeneous pool of cells harboring targeted disruptions in the DNAJB14 gene, generated by CRISPR/Cas9-mediated gene editing. This population-level knockout approach facilitates studies where clonal variability is minimized, and bulk cellular responses to DNAJB14 ablation are of primary interest.
The NCI-H1299 cell line originates from a lymph node metastasis of a lung adenocarcinoma and is widely used as a model for non-small cell lung carcinoma (NSCLC) and metastasis. These cells are TP53 null and express wild-type EGFR, which are characteristic genetic features of many lung adenocarcinomas. NCI-H1299 cells exhibit epithelial morphology and have been extensively employed in cancer biology to study tumor progression, drug resistance, and protein homeostasis mechanisms. The absence of functional p53 in these cells heightens reliance on chaperone networks for survival under proteotoxic stress, making this host particularly relevant for examining co-chaperone functions.
DNAJB14 encodes a J-domain co-chaperone that stimulates Hsp70 ATPase activity, facilitating substrate binding, folding, and degradation. It interacts with HSPA1A/Hsp70, other DnaJ family members, and BAG co-chaperones such as BAG3 to coordinate protein triage between refolding and degradation. DNAJB14 is transcriptionally regulated by HSF1 and induced under endoplasmic reticulum and oxidative stress, linking it to the unfolded protein response. Downstream, it promotes client protein folding and modulates the ubiquitin-proteasome pathway; key components include HSP90, BAG3, and STUB1/CHIP. By regulating Hsp70 ATPase activity, DNAJB14 maintains proteostasis, and its disruption shifts the balance toward proteotoxic stress and altered protein degradation.
In the NCI-H1299 lung adenocarcinoma background, knockout of DNAJB14 is predicted to compromise protein quality control, potentially exacerbating proteotoxic stress and impacting cancer cell fitness. Given the TP53-null status and reliance on stress response pathways, these polyclonal knockout cells offer a platform to dissect how co-chaperone dysfunction affects tumor cell survival, particularly under conditions that challenge the chaperone network, such as chemotherapy or targeted therapy. The model enables interrogation of the interplay between DNAJB14, Hsp70, and downstream ubiquitin-proteasome activity in an established lung cancer metastasis context, providing insights relevant to both cancer biology and protein misfolding disorders.
This model supports investigation of chaperone function in cancer, protein homeostasis, and drug resistance. Common endpoints include expression analysis via Western blot and RT-qPCR, protein interaction studies by co-immunoprecipitation, cell viability assays, proteasome activity measurements, and confocal microscopy for stress granule dynamics. The polyclonal population is suitable for high-content screens and functional complementation experiments, and can be used to test chaperone-targeted therapeutics or evaluate dependency on DNAJB14-mediated proteostasis. For additional details, please contact Ascent Research.
