The DNAJC10 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population with targeted disruption of the DNAJC10 gene in the 786-O renal cell carcinoma line. This knockout model provides a heterogeneous loss-of-function system, avoiding clonal artifacts and enabling robust population-level analysis of DNAJC10 function.
The 786-O line, derived from a human primary clear cell renal adenocarcinoma, is a well-established model for clear cell renal cell carcinoma (ccRCC) and kidney epithelial tumorigenesis. Its stable phenotype and reproducible growth characteristics make it an ideal host for CRISPR/Cas9-mediated gene editing, enabling robust comparative studies between knockout and parental populations.
DNAJC10 encodes an ER luminal J-domain co-chaperone that enhances Hsp70 ATPase activity to facilitate protein folding and assembly. It functions within the unfolded protein response (UPR), acting downstream of ER stress sensors IRE1, PERK, and ATF6, which are triggered by conditions like hypoxia or nutrient deprivation. DNAJC10 interacts with BiP/GRP78 and Hsp70 to mitigate proteotoxic stress; CRISPR-mediated disruption of this gene impairs the adaptive UPR, leading to sustained PERK and IRE1 signaling, increased expression of the pro-apoptotic transcription factor CHOP and GADD34, activation of JNK, and ultimately sensitization to ER stress-induced apoptosis.
In 786-O cells, which experience heightened basal ER stress due to their tumorigenic nature, DNAJC10 provides crucial cytoprotection. CRISPR-mediated knockout abrogates this protection, disrupting the adaptive UPR balance and promoting apoptosis through CHOP and JNK pathways. The resulting sensitization to ER stress inducers such as tunicamycin or proteasome inhibitors makes this model valuable for studying renal cancer cell vulnerabilities. The polyclonal composition reflects tumor heterogeneity, yielding population-level insights into chaperone dysfunction.
Typical applications include detailed UPR pathway analysis via Western blotting for BiP and CHOP, RT-qPCR for XBP1 mRNA splicing, and apoptosis detection through caspase-3 cleavage assays. The cells can be subjected to tunicamycin-induced ER stress for viability assays, drug sensitivity testing with proteasome inhibitors, and migration/invasion studies. Immunofluorescence for ER stress granules further allows spatial monitoring of stress responses. This polyclonal knockout model is ideal for research in renal cell carcinoma biology, ER stress-related diseases, and the development of chaperone-targeted therapeutics. For additional technical information, contact Ascent Research.
