The HSPBP1 Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of HSPBP1 in the HAP1 human near-haploid leukemia cell line. This loss-of-function model enables investigation of co-chaperone HSPBP1??s role in protein homeostasis and apoptosis. The polyclonal pool contains heterogeneous edited cells for robust population-level studies, supplied as frozen pellets for immediate culture expansion.
The HAP1 cell line is a human near-haploid leukemia cell line derived from the KBM-7 chronic myeloid leukemia patient line. These cells are adherent and maintain a near-haploid karyotype for most chromosomes, making them an ideal model system for functional genomics and CRISPR-based gene disruption studies. The haploid genetic background simplifies genetic analysis and facilitates efficient knockout generation, as only a single allele typically requires disruption. HAP1 cells retain key signaling pathways relevant to cancer biology, including stress responses and apoptotic machinery.
HSPBP1 encodes a co-chaperone that regulates HSP70 ATPase activity, acting as a nucleotide exchange factor to promote ADP release and substrate dissociation. Under stress such as heat shock or oxidative stress, its expression is upregulated by HSF1. HSPBP1 interacts with HSP70, HSPA8, BAG proteins, STUB1/CHIP, and STIP1/HOP, forming complexes that dictate client protein fate??refolding, degradation, or aggregation. By inhibiting HSP70 ATPase, HSPBP1 can suppress apoptosis via modulation of caspase activation and mitochondrial integrity, stabilizing anti-apoptotic clients and aiding misfolded protein clearance.
Disruption of HSPBP1 in the HAP1 near-haploid background offers a clean genetic model to dissect chaperone-mediated quality control and apoptosis regulation. Functional stress pathways in HAP1 cells allow examination of HSPBP1-dependent effects on HSP70 activity, client turnover, and sensitivity to proteotoxic insults. Given HSPBP1??s roles in cancer survival, neurodegeneration, and ischemic injury, this model aids studies of chemoresistance, protein aggregation, and stress-induced death.
These polyclonal knockout cells support western blotting, co-immunoprecipitation, and ATPase assays to evaluate HSP70 regulation. Apoptosis can be monitored via caspase-3 activation and Annexin V staining. Applications extend to fluorescence microscopy for protein aggregation, RT-qPCR, proteasomal activity assays, and drug sensitivity tests with proteasome or HSP70 inhibitors. The model is suited for functional screens and chaperone-targeted therapy research. For details, contact Ascent Research.
