The HSPA4 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population that provides stable loss of HSPA4 function in the HEK293T background. This product consists of a heterogeneous pool of cells carrying targeted gene disruptions, enabling bulk analysis of HSPA4-dependent processes without clonal selection artifacts. The knockout is achieved via CRISPR/Cas9-mediated gene disruption, and the polyclonal format captures diverse editing outcomes to ensure robust population-level depletion of the HSPA4 chaperone.
HEK293T cells are human embryonic kidney epithelial cells transformed with adenovirus type 5 DNA and stably expressing the SV40 large T antigen. Derived from the HEK293 line, this host exhibits high transfection efficiency and supports episomal replication of plasmids containing the SV40 origin, making it a standard platform for transient protein overexpression, lentiviral packaging, and gene editing. The fast growth and adaptability to high-throughput formats render HEK293T ideal for generating knockout models that require consistent cell sources for downstream assays.
HSPA4 is an ATP-dependent molecular chaperone of the HSP70 family that maintains proteostasis by refolding misfolded proteins or targeting them for ubiquitin-mediated degradation. Its expression is induced by HSF1 and HSF2 in response to hyperthermia, oxidative stress, and DNA damage. HSPA4 cooperates with co-chaperones HSP40 (DNAJ family), HSP90, and BAG proteins (e.g., BAG3) and the E3 ligase STUB1/CHIP to manage client proteins. Additionally, HSPA4 directly binds Apaf-1 to inhibit apoptosome formation and sequesters AIF to prevent caspase-independent apoptosis, thereby linking chaperone activity to regulation of BAX, Bcl-2, and NF-??B-mediated survival signaling.
In the HEK293T context, HSPA4 knockout allows dissection of the cross-talk between proteostasis and apoptotic pathways. The cells’ high transfectability enables reintroduction of wild-type or mutant HSPA4 along with client proteins to map functional domains. Population-level assays such as Western blotting and co-immunoprecipitation can probe endogenous interactions and downstream target expression. Furthermore, the knockout sensitizes cells to stress-induced apoptosis, providing a model to test compounds that target chaperone addiction in cancer or to evaluate neuroprotective interventions that modulate apoptotic thresholds.
Typical applications include heat shock and oxidative stress assays with HSF1 luciferase reporter readouts, cell viability (MTT/CCK-8) and Annexin V apoptosis measurements, and co-immunoprecipitation to verify interactions with HSP40, HSP90, BAG3, STUB1, or Apaf-1. The model supports cancer therapy evaluation as a negative-control background for HSPA4 inhibitors and can be used in neurodegenerative disease studies to assess chaperone-mediated suppression of protein aggregation. For more details, contact Ascent Research.
