The DNAJC3 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the human 786-O renal cell carcinoma line, featuring targeted disruption of the DNAJC3 gene. This knockout model provides a robust tool for investigating the roles of DNAJC3/p58IPK in the unfolded protein response (UPR), PKR signaling, and ER stress biology. The polyclonal format ensures a heterogeneous knockout population, enabling functional studies that reflect varied gene-editing outcomes without clonal selection bias.
The 786-O host cell line is an established model of clear cell renal cell carcinoma (ccRCC), a common and aggressive kidney cancer subtype. These adherent epithelial cells harbor characteristic VHL mutations that mimic the genetic landscape of ccRCC, making them highly relevant for studying tumor cell signaling, metabolic adaptation, and therapeutic vulnerabilities. Their use as the host for DNAJC3 knockout facilitates the investigation of UPR and ER stress pathways specifically within the context of kidney cancer pathogenesis.
DNAJC3 encodes the co-chaperone p58IPK, which functions as an inhibitor of the dsRNA-activated protein kinase PKR and a negative regulator of the unfolded protein response (UPR). p58IPK interacts with PERK, IRE1, HSP70, and GRP78/BiP to attenuate eIF2?? phosphorylation and downstream ATF4?CCHOP signaling, while also suppressing PKR-mediated activation of JNK and NF-??B. Knockout of DNAJC3 removes this inhibitory control, leading to unchecked PKR activity, sustained eIF2?? phosphorylation, and potentiated ER stress-induced apoptosis via CHOP and caspase cascades.
In the 786-O ccRCC background, loss of DNAJC3 function is significant because kidney cancer cells often face ER stress from rapid proliferation, hypoxia, and metabolic dysregulation. The exacerbated UPR and PKR signaling resulting from DNAJC3 knockout may heighten apoptosis susceptibility and alter pro-survival signaling, offering a valuable model to dissect how UPR modulation impacts ccRCC cell fate and to screen for ER stress-inducing therapeutic agents.
Researchers can utilize these DNAJC3 knockout polyclonal cells for mechanistic studies of ER stress and UPR signaling, functional analysis in kidney cancer, drug sensitivity profiling under ER stress inducers such as tunicamycin, and elucidation of PKR-mediated apoptosis mechanisms. Typical assays include western blotting for BiP and CHOP, phospho-eIF2?? immunoblotting, cell viability and caspase-3/7 activation assays, PKR kinase activity measurements, and colony formation assays. For further information, please contact Ascent Research.
