The DNAJB2 Knockout 786-O Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population of human 786-O renal carcinoma cells carrying heterogeneous mutations in the DNAJB2 gene. This mixed pool, generated by non?homologous end joining, provides a loss-of-function model that avoids clonal artifacts and captures the full spectrum of gene-disruption effects.
The parental 786-O line is a clear cell renal cell carcinoma (ccRCC) isolate characterized by a biallelic VHL frameshift mutation. This genetic lesion stabilizes hypoxia-inducible transcription factors, rewires metabolism, and imposes chronic proteotoxic stress, thereby creating a highly relevant background for studying co-chaperone networks and protein quality-control mechanisms.
DNAJB2 functions as an Hsp40 co-chaperone that partners with Hsp70 to triage misfolded proteins. Under stress conditions, HSF1 transcriptionally upregulates DNAJB2, which then recruits Hsp70 to aberrant substrates. Through interactions with BAG1, BAG2, and the E3 ligase STUB1/CHIP, DNAJB2 stimulates Hsp70 ATP hydrolysis, promoting either client refolding or ubiquitin-dependent proteasomal degradation. This molecular switch determines cellular proteostasis outcomes.
In VHL?mutant 786-O cells, DNAJB2 disruption accentuates proteotoxic imbalances, revealing the co-chaperone??s role in sustaining ccRCC viability. The knockout model enables dissection of how the DNAJB2?CHsp70?CCHIP axis governs the fate of misfolded oncogenic or tumor?suppressor proteins, and facilitates identification of synthetic lethal interactions with proteasome or Hsp70 inhibitors.
Representative applications for this polyclonal knockout product include western blotting for ubiquitinated protein accumulation, proteasome activity assays, detergent?insoluble protein aggregation measurements, and cell viability assessments under heat shock or oxidative stress. Co?immunoprecipitation studies reveal altered chaperone?Cclient complexes. The model supports research into distal hereditary motor neuropathy, Charcot?Marie?Tooth disease type 2, and ccRCC stress responses. For further information, contact Ascent Research.
