The DNAJC1 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the 786-O human renal cell carcinoma line, designed for loss-of-function analysis of DNAJC1. This heterogeneous pool of gene-disrupted cells avoids clonal selection biases and provides a robust model for studying DNAJC1 function in cancer biology. The polyclonal format facilitates population-level assays such as drug sensitivity screening and transcriptomic profiling, enabling dissection of DNAJC1??s role in chaperone-mediated protein homeostasis and oncogenic signaling.
The 786-O cell line is a well-characterized model of clear cell renal cell carcinoma (ccRCC), originally established from a primary tumor of a 58-year-old male. These VHL-mutant cells exhibit constitutive HIF1A activation, driving tumorigenic programs such as angiogenesis and metabolic reprogramming. As proximal tubular epithelial cancer cells, 786-O retains kidney-specific features, making it an ideal system to study renal carcinoma biology, particularly ER stress and chaperone function.
DNAJC1 encodes an Hsp40 co-chaperone that partners with Hsp70 (HSPA1A) and Hsc70 (HSPA8) to stimulate ATPase activity, facilitating client protein folding and degradation. It is critical for the unfolded protein response (UPR) and ER stress recovery. DNAJC1 expression is regulated by HSF1, ATF4, XBP1, p53, HIF1A, and MYC. It influences downstream AKT and ERK1/2 signaling and modulates BCL-2 family members (BCL-2, BAX), integrating chaperone activity with survival and apoptosis. DNAJC1 interacts with BAG-1, HOP, CHIP, and GRP78/BiP, positioning it at a key signaling hub.
Disruption of DNAJC1 in VHL-mutant 786-O cells impairs Hsp70 chaperone cycle activity, causing misfolded protein accumulation and unresolved ER stress. This activates UPR sensors PERK, IRE1, and ATF6, leading to CHOP induction and p53 activation. Consequently, cells undergo p53-dependent apoptosis and reduced proliferation due to diminished ERK1/2 phosphorylation and altered CDK4/Cyclin D1 expression. The polyclonal model enables analysis of heterogeneous stress responses and drug sensitivities, making it valuable for preclinical studies targeting the Hsp70 system or UPR in renal cancer.
These polyclonal knockout cells support a variety of functional assays. Western blotting can assess DNAJC1, Hsp70, p53, and phospho-ERK1/2 levels; RT-qPCR quantifies UPR markers such as CHOP and GRP78. Apoptosis is detected by Annexin V/PI staining, and proliferation by MTT or BrdU incorporation. Co-immunoprecipitation enables mapping of Hsp70 client proteins and interactions. The cells are also ideal for drug sensitivity profiling with proteasome inhibitors (e.g., bortezomib) or Hsp inhibitors, and transcriptome-wide analysis via RNA-seq. For ordering information, please contact Ascent Research.
