The HSPH1 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in the HeLa background, designed for targeted disruption of the HSPH1 gene. This heterogeneous pool of gene-edited cells provides a loss-of-function model for investigating HSPH1-dependent processes without clonal selection, making it suitable for studying population-level stress responses and chaperone network dynamics.
HeLa cells are an extensively characterized human epithelial cell line derived from a cervical adenocarcinoma originally isolated from Henrietta Lacks. As a widely employed model in cancer biology, molecular biology, and drug discovery, HeLa cells offer robust growth, ease of manipulation, and a well-documented genetic background. Their tumorigenic origin makes them particularly relevant for examining oncogenic stress adaptation and therapy resistance mechanisms.
HSPH1 (Hsp105) functions as a nucleotide exchange factor for HSP70, accelerating ADP release and ATP binding to enhance HSP70-mediated protein folding and disaggregation. HSPH1 is transcriptionally upregulated by HSF1 in response to heat shock, oxidative stress, and proteotoxic stress, and it interacts directly with HSP70, Hsp40 (DNAJ proteins), Hsp90, and apoptosis-inducing factor (AIF). Within the heat shock response, ER stress response, and unfolded protein response pathways, HSPH1 promotes solubilization of protein aggregates and refolding of denatured clients; its knockout impairs cellular thermotolerance, increases protein aggregation, and reduces viability under stress conditions.
In the HeLa cervical adenocarcinoma model, HSPH1 disruption enables dissection of chaperone-dependent stress adaptation critical for cancer cell survival. HeLa cells rely on robust protein quality control to cope with oncogenic proteotoxicity, and loss of HSPH1 sensitizes them to proteasome inhibition and chemotherapeutic agents. This knockout system thus provides a platform to study how nucleotide exchange factor activity modulates aggregate clearance and stress signaling in a relevant cancer background.
These polyclonal knockout cells support diverse experimental applications, including assessment of protein folding and aggregation via Western blotting for HSPH1 and HSP70, RT-qPCR for HSPH1 transcript levels, and heat shock induction. Co-immunoprecipitation assays can probe HSPH1-HSP70 complex formation, while cell viability assays under oxidative or chemotherapeutic stress quantify functional consequences. Immunofluorescence staining for ubiquitinated aggregates and flow cytometry for apoptosis after stress further characterize the role of HSPH1 in chaperone-mediated cytoprotection. The model is applicable to research on cancer cell stress biology, neurodegenerative protein aggregation, and ischemia-reperfusion injury. For further information or technical support, please contact Ascent Research.
