The DNAJB2 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 human hepatocellular carcinoma line. This loss-of-function model enables targeted disruption of the DNAJB2 gene without clonal isolation, offering a genetically diverse pool for studying co-chaperone biology. The CRISPR/Cas9-mediated gene disruption eliminates DNAJB2 expression, facilitating interrogation of its role in proteostasis.
The Huh-7 host cell line is a well-differentiated, epithelial hepatocellular carcinoma model extensively used to investigate hepatic metabolism, protein synthesis, and liver cancer biology. Its robust protein folding machinery and active secretory pathways provide a relevant cellular environment for analyzing chaperone network dynamics.
DNAJB2 (Hsp40 homolog) is a molecular co-chaperone that recognizes misfolded proteins and recruits HSPA1A/Hsp70. Through stimulation of Hsp70 ATPase activity, it promotes substrate handover to the ubiquitin ligase STUB1/CHIP, which collaborates with BAG-1 to direct clients toward proteasomal degradation. DNAJB2 expression is induced by the transcription factor HSF1 under conditions of heat shock or oxidative stress, linking it to the unfolded protein response and chaperone-mediated autophagy. This gene is therefore pivotal in clearing aggregation-prone proteins and preserving proteostasis.
In hepatocellular carcinoma cells, DNAJB2 knockout compromises the proteostatic buffer, potentially sensitizing cells to stress-induced apoptosis and protein aggregation. Given that proteotoxic stress and chaperone imbalances are implicated in liver tumorigenesis, this model allows dissection of DNAJB2 contributions to cancer cell survival. Furthermore, DNAJB2 mutations cause spinocerebellar ataxia, highlighting its importance in neuronal protein quality control; the Huh-7 knockout system thus offers a unique human-derived platform to study neurodegenerative mechanisms within an epithelial cancer background.
Typical experimental uses include protein aggregation assays, proteasome activity measurements, co-immunoprecipitation of chaperone complexes, and Western blot or RT-qPCR analysis of downstream effectors. Additional applications encompass immunofluorescence-based detection of aggregate formation, apoptosis assays to assess stress sensitivity, and screening for modulators of the HSF1?CHsp70?CDNAJB2 pathway. This polyclonal knockout model is a valuable tool for both basic proteostasis research and translational studies in cancer and neurodegeneration. For product inquiries, contact Ascent Research.
