The DNAJB2 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the DNAJB2 gene has been disrupted in the HEK293T human embryonic kidney cell background. This product provides a heterogeneous pool of edited cells that facilitates loss-of-function studies of DNAJB2, encoding a co-chaperone critical for protein quality control. The polyclonal format avoids clonal selection artifacts and offers a robust model for interrogating chaperone-mediated processes without the confounding effects of single-cell bottlenecks.
HEK293T cells, derived from human embryonic kidney 293 cells stably expressing the SV40 large T antigen, are an adherent epithelial cell line widely employed for protein expression, lentiviral and retroviral packaging, and mechanistic investigations. Their high transfection efficiency and robust protein production make them an ideal host for creating knockout populations aimed at dissecting cellular pathways in a genetically tractable system. The HEK293T background enables seamless integration of the DNAJB2 knockout into existing workflows for protein homeostasis and neurobiology research.
DNAJB2 functions as a co-chaperone that interacts directly with Hsc70 (HSPA8) to modulate protein folding, refolding, and degradation. It recruits misfolded substrates to Hsc70 and, in concert with ubiquitin, BAG family proteins, and 26S proteasome subunits, tags clients for proteasomal destruction. DNAJB2 also participates in chaperone-mediated autophagy by facilitating substrate translocation through LAMP2A. Its expression is regulated by heat shock factor 1 (HSF1) and cellular stress signals, while its activity impacts the unfolded protein response and ER-associated degradation pathways. Loss of DNAJB2 disrupts this network, leading to accumulation of aggregation-prone proteins.
In the HEK293T context, depletion of DNAJB2 creates a model for studying proteostasis failure and its downstream neurotoxic consequences. This knockout is directly relevant to peripheral neuropathies, including Charcot-Marie-Tooth disease type 2T and distal hereditary motor neuropathy, where DNAJB2 mutations compromise neuronal protein quality control. The cell population allows researchers to examine how impaired co-chaperone function affects ubiquitin-proteasome system activity, autophagy flux, and cellular vulnerability to proteotoxic stress, providing insights into the molecular underpinnings of neurodegeneration.
Researchers can employ this polyclonal knockout model in diverse applications, including western blotting and co-immunoprecipitation to assess DNAJB2 and HSPA8 interactions, proteasome activity assays to measure degradation capacity, and immunofluorescence staining to visualize protein aggregates. Cell viability assays under proteotoxic insult, autophagy flux reporters, and flow cytometry for apoptotic markers further enable detailed phenotypic characterization. Small-molecule screens aimed at enhancing proteostasis or restoring protein clearance can also be conducted. For further information, please contact Ascent Research.
