The HSPH1 Knockout HEK293T Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human HSPH1 gene in HEK293T cells. This loss-of-function model is designed to investigate heat shock response and chaperone-mediated protein folding. The polyclonal pool offers heterogeneous gene disruption, enabling robust functional studies without clonal selection biases. It is suitable for pooled screening, drug response assays, and proteotoxic stress analysis.
The host line, HEK293T, is a human embryonic kidney epithelial cell line immortalized with adenovirus 5 DNA and expressing SV40 large T antigen. These characteristics confer high transfection efficiency, strong protein expression, and effective viral packaging capabilities, making it a standard system for biochemical and pharmacological research. The epithelial phenotype and rapid growth facilitate reproducible, scalable experiments. In the HSPH1 knockout context, HEK293T provides a well-characterized background for studying chaperone network function under stress.
HSPH1 encodes Hsp110, a stress-inducible molecular chaperone that acts as a nucleotide exchange factor for Hsp70. Under stress conditions such as hyperthermia, the transcription factor HSF1 upregulates HSPH1 expression. Hsp110 interacts directly with Hsp70, Hsp40, STIP1, and BAG family proteins to promote ADP release and ATP binding, driving client protein refolding or degradation. Key clients include mutant p53 and tau, linking the HSF1??HSPH1??Hsp70 axis to proteostasis, apoptosis, and neurodegeneration. This pathway is critical for maintaining protein solubility and cellular viability during proteotoxic challenges.
In HEK293T cells, HSPH1 knockout impairs the stress-responsive chaperone network, sensitizing cells to heat shock, oxidative stress, and protein misfolding. This model enables dissection of Hsp110’s role in a human epithelial cellular environment, particularly relevant given HEK293T’s wide use in mechanistic and drug discovery studies. The polyclonal nature helps capture the functional heterogeneity often observed in tumor cell populations, facilitating research into cancer cell stress resilience, synthetic lethality, and the impact of chaperone loss on client protein stability.
Typical applications include proteostasis studies, Hsp110 inhibitor screening, and chaperone interaction analysis. Researchers can employ western blotting to monitor HSPH1 and Hsp70 expression, viability assays following heat shock, protein aggregation assessments, apoptosis detection, and co-immunoprecipitation to examine Hsp70 complexes. The model also supports high-content imaging and genetic modifier screens. For additional information, please contact Ascent Research.
