This product comprises a CRISPR/Cas9-edited polyclonal HEK293T cell population with targeted disruption of the BAG2 gene. The cells are supplied as a heterogeneous knockout pool, each carrying distinct gene-editing events. The polyclonal design avoids clonal artefacts and provides a robust loss-of-function model for studying BAG2-dependent processes, eliminating interference from endogenous wild-type protein.
The HEK293T host cell line derives from human embryonic kidney HEK293 cells and stably expresses the SV40 large T antigen. This enables episomal plasmid replication and leads to high-level recombinant protein production and efficient viral packaging. The cells exhibit an epithelial morphology, rapid adherent growth, and outstanding transfectability, making them ideal for knockout model generation and downstream overexpression or complementation assays.
BAG2 encodes a co-chaperone that binds the ATPase domain of HSP70/HSC70, facilitating release of misfolded substrates for 26S proteasomal degradation. It competes with the STUB1 (CHIP) ubiquitin ligase, thereby inhibiting substrate ubiquitination and modulating the balance between degradation and aggregation. BAG2 is regulated by heat shock, ER stress, and proteasome inhibition, and its downstream effects include stabilization of anti-apoptotic factors such as BAX. Direct interaction with proteasome subunits further positions BAG2 at a key proteostasis checkpoint.
Disrupting BAG2 in HEK293T cells creates a sensitive system to delineate proteostasis networks, as the cell line??s robust protein expression machinery heightens the load of misfolded polypeptides. The polyclonal knockout pool permits bulk biochemical studies while averaging out editing heterogeneity, thus reflecting population-level responses. This model reveals phenotypes linked to aggregate accumulation, altered proteasome flux, and apoptotic signaling perturbations, providing a physiologically relevant context compared to clonal lines.
Applications include dissecting HSP70-mediated triage, ubiquitin-proteasome function, and apoptosis control. The cells are used to model neurodegenerative proteotoxicity via overexpression of aggregation-prone proteins, to investigate chemoresistance mechanisms in cancer, and to screen for modulators of protein clearance. Compatible assays include Western blotting, RT-qPCR, proteasome activity measurements, ubiquitination assays, immunofluorescence for aggregates, co-immunoprecipitation of HSP70 complexes, and caspase-3 activation assays. Contact Ascent Research for further information or custom applications.
