The BAG3 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated by disruption of the BAG3 gene in the human embryonic kidney cell line HEK293T. This loss-of-function model is designed for researchers investigating BAG3-dependent pathways in autophagy, apoptosis, and cellular stress responses. The polyclonal format provides a heterogeneous pool of knockout cells, enabling robust assessment of BAG3 function without clonal bias.
HEK293T cells are a derivative of the HEK293 human embryonic kidney cell line that stably expresses the SV40 large T antigen, conferring high episomal replication and protein expression capabilities. These adherent epithelial cells are widely employed for recombinant protein production, lentiviral and retroviral packaging, and transient gene expression studies. The robust growth and high transfection efficiency of HEK293T make it an ideal host for creating knockout models to investigate fundamental cellular processes.
BAG3 encodes a co-chaperone that orchestrates chaperone-assisted selective autophagy (CASA), a pathway responsible for lysosomal clearance of aggregation-prone and damaged proteins. BAG3 forms complexes with HSPA8/Hsc70 and HSPB8, recruiting them to ubiquitinated substrates and linking them to the autophagy machinery through direct interaction with p62/SQSTM1 and LC3. Upstream, BAG3 transcription is induced by HSF1 and NF-??B in response to heat shock, oxidative stress, and proteasome inhibition. Downstream, BAG3 modulates apoptosis by interacting with Bcl-2 family proteins and influences cytoskeletal dynamics through vimentin and filamin. Additional interacting partners such as 14-3-3 proteins and phospholipase C-gamma highlight its integration into diverse stress and survival signaling cascades.
Disruption of BAG3 in the HEK293T background provides a physiologically relevant system for examining the functional consequences of impaired CASA and altered proteostasis. Given the endogenous expression of key autophagy and apoptosis regulators in HEK293T cells, this knockout model enables precise dissection of BAG3-dependent processes such as protein aggregate clearance, stress-induced cell signaling, and cell death regulation. Moreover, the transformed phenotype of HEK293T cells renders this model particularly valuable for cancer biology research, as BAG3 is known to support tumor cell survival and chemoresistance in glioblastoma, pancreatic, and breast cancers.
Typical downstream applications include monitoring autophagy flux via LC3 turnover and p62 accumulation assays, assessing apoptosis by Annexin V/PI staining, and evaluating cell viability under conditions of serum starvation or proteasome inhibitor treatment. Co-immunoprecipitation experiments can be used to validate interactions with HSPA8, HSPB8, or p62, while RT-qPCR permits quantification of stress-responsive transcriptional programs. The model is also suited for drug screening efforts aimed at identifying small-molecule modulators of BAG3 or CASA activity. Additionally, researchers studying myofibrillar myopathies and dilated cardiomyopathy will find these cells useful for exploring BAG3??s role in muscle proteostasis. For additional product information or technical assistance, please contact Ascent Research.
