The HSPB8 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-disrupted polyclonal knockout population of the HSPB8 gene in HEK293T cells, providing a heterogeneous loss-of-function model to study small heat shock protein B8-dependent processes. This polyclonal format reduces clonal biases and is suitable for bulk assays investigating autophagy, stress response, and proteostasis in a transferable and widely used host background.
HEK293T is an immortalized human embryonic kidney epithelial cell line stably expressing the SV40 large T antigen, known for high transfectability and episomal plasmid replication, supporting robust recombinant protein expression and lentiviral production. These adherent cells are permissive for viral replication and offer a clean background for dissecting signaling pathways relevant to human disease.
HSPB8 is a co-chaperone that mediates chaperone-assisted selective autophagy (CASA) by interacting with BAG3, Hsc70, and STUB1 to target ubiquitinated misfolded proteins to autophagosomes via p62/SQSTM1 and LC3. This process is activated by heat shock and oxidative stress downstream of HSF1 and TNF-alpha signaling, and it prevents apoptosis by regulating caspase activation while maintaining cytoskeletal integrity. Disruption of HSPB8 impairs proteostasis and autophagic flux, modeling defects seen in Charcot-Marie-Tooth disease type 2L and related myopathies.
In HEK293T cells, HSPB8 knockout provides a tractable system to study CASA machinery under induced stress, examining how loss of this small heat shock protein impacts aggregate clearance and cell survival. The immortalized nature permits extensive genetic and pharmacological manipulation, making it valuable for investigating neurodegeneration and muscle disease mechanisms where protein aggregation is central.
Representative assays include Western blotting for autophagy markers, LC3-II turnover for autophagy flux, immunofluorescence for aggregate detection, caspase-3 activation assays, and RT-qPCR. Applications range from drug screening for proteostasis modulators to mechanistic studies of peripheral neuropathy and myofibrillar myopathy. For further inquiries, please contact Ascent Research.
