The DNAJB4 Knockout HEK293T Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population derived from HEK293T human embryonic kidney cells. This product targets the DNAJB4 gene in Homo sapiens, generating a heterogeneous pool of alleles with disrupted gene function. The polyclonal format provides a research-ready tool for studying gene loss-of-function without clonal isolation, enabling robust analysis of protein quality control pathways.
The HEK293T host cell line is immortalized with SV40 large T antigen, facilitating high-level transient transfection, protein expression, and viral production. This cell line is a widely used platform for investigating signaling mechanisms and stress responses, making it an ideal model for CRISPR-mediated knockout studies of chaperone genes. Its rapid proliferation and manipulability support advanced cellular assays in cancer biology and neurodegeneration research.
DNAJB4 functions as an Hsp40 co-chaperone that recruits Hsp70 (HSPA1A) to misfolded proteins, promoting their refolding or ubiquitin-dependent proteasomal degradation. Its expression is activated by HSF1 under conditions of heat shock, oxidative stress, and proteotoxic stress. DNAJB4 interacts with BAG3, STUB1/CHIP, HSP90, and HSP110, integrating the unfolded protein response, chaperone-mediated autophagy, and the ubiquitin-proteasome system. Disruption of DNAJB4 impairs the processing of Hsp70 client proteins and increases the burden of aggregated, misfolded species.
Loss of DNAJB4 in HEK293T cells compromises cellular proteostasis, heightening sensitivity to proteotoxic agents. This knockout model is valuable for examining protein aggregation diseases, including neurodegenerative conditions like Alzheimer??s and Parkinson??s, and cancers where chaperone networks are often deregulated. The polyclonal population enables study of stress response dynamics without clonal bias, providing insights into the roles of co-chaperones in disease-relevant contexts.
These cells support diverse assays, such as co-immunoprecipitation with Hsp70 to assess substrate interactions, ubiquitination assays to monitor proteasomal targeting, and proteasome activity measurements. Protein aggregation can be tracked using biochemical or imaging methods. Cell viability under stress, western blotting for HSPA1A and BAG3, and RT-qPCR for chaperone target genes allow comprehensive profiling of the proteostasis network. Applications extend to cancer biology and neurodegeneration modeling. For further details, please contact Ascent Research.
