The DNAJB1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HEK293T cells, designed for loss-of-function studies of the DNAJB1 gene. This heterogeneous pool is generated through CRISPR/Cas9-mediated gene disruption without clonal isolation, capturing a broad spectrum of target-gene mutations and providing a robust model for pooled functional assays and high-throughput screening applications while avoiding clonal selection bias.
The HEK293T host cell line is derived from human embryonic kidney cells immortalized with adenovirus 5 E1A/E1B DNA and stably expressing the SV40 large T-antigen. These cells display epithelial morphology with neuronal lineage gene expression patterns and are one of the most widely employed mammalian expression systems, valued for high transfection efficiency, robust protein production, and compatibility with viral packaging and recombinant protein expression platforms.
DNAJB1 encodes a member of the Hsp40 co-chaperone family that serves as an essential regulator of Hsp70. It recruits Hsp70 to client proteins and stimulates its ATP hydrolysis, thereby driving protein folding, refolding, or ubiquitin-proteasome-mediated degradation. Transcription of DNAJB1 is up-regulated by HSF1 upon cellular stress such as heat shock, oxidative damage, or proteotoxic insults. Downstream, the DNAJB1?CHsp70 complex modulates apoptosis through JNK and p53 pathways and affects NF-??B signaling. Key interacting partners include Hsp90, BAG family co-chaperones, CHIP (STUB1), and tau, which together coordinate protein triage decisions between folding and degradation.
In the HEK293T context, loss of DNAJB1 provides a valuable tool for dissecting stress-responsive signaling networks and proteostasis maintenance. Given the cell line’s active protein synthesis and the influence of SV40 T-antigen on cell cycle control, DNAJB1 knockout may sensitize cells to heat shock, impair the unfolded protein response, and alter the handling of aggregation-prone proteins. This enhances the model’s utility for studying protein misfolding diseases such as Parkinson??s and Alzheimer??s, as well as cancers including hepatocellular carcinoma and renal cell carcinoma, where chaperone dysregulation is implicated.
These polyclonal knockout cells support a wide range of experimental applications in chaperone biology, stress response, and apoptosis research. Researchers can confirm target disruption by western blotting for DNAJB1 and Hsp70, assess chaperone?Cclient binding via co-immunoprecipitation, measure HSF1 activity using luciferase reporters, evaluate protein thermal stability, detect apoptosis with Annexin V/PI staining, monitor proteasome activity, and visualize protein aggregation by immunofluorescence. Additional assays include heat shock survival testing and drug sensitivity screening. For further information, please contact Ascent Research.
