The DNAJB5 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the HeLa cell line, engineered to disrupt the expression of the DNAJB5 gene. This product provides a heterogeneous pool of knockout cells generated through CRISPR/Cas9-mediated gene targeting, offering a robust loss-of-function model for investigating the molecular functions of DNAJB5 without clonal selection. The polyclonal format preserves genetic diversity while ensuring efficient target-gene disruption, making it suitable for functional genomics studies and screening applications where monoclonal isolation is not required.
HeLa cells are a widely used immortalized human cell line originally derived from a cervical adenocarcinoma. This HPV18-positive line exhibits inactivation of the p53 and retinoblastoma (Rb) tumor suppressor pathways, contributing to its robust proliferation and transformation phenotype. HeLa cells serve as a standard model in cancer biology, protein expression, and functional genomics due to their ease of culture, well-characterized genome, and susceptibility to gene editing. The integration of a DNAJB5 knockout into this background enables the study of chaperone function within the context of HPV-driven oncogenesis and cellular stress adaptation.
DNAJB5 belongs to the DNAJ/Hsp40 family of co-chaperones and functions as a critical regulator of the heat shock response and protein quality control. It interacts directly with Hsp70 through its J-domain, stimulating ATP hydrolysis to promote substrate binding and facilitate refolding or degradation of misfolded and aggregated proteins. DNAJB5 is transcriptionally activated by HSF1 under thermal or ER stress, and it cooperates with Hsp90, CHIP, and HOP to coordinate protein triage between refolding and degradation. Moreover, DNAJB5 participates in clathrin-mediated endocytosis by binding clathrin, linking chaperone function to membrane trafficking. Disruption of DNAJB5 thus compromises proteotoxic stress management, leading to protein aggregate accumulation and altered endocytic dynamics.
In the HeLa cervical cancer background, knockout of DNAJB5 provides a powerful platform to dissect chaperone networks under conditions of constitutive oncogenic stress. The loss of this co-chaperone may exacerbate protein aggregation and sensitize cells to proteasome inhibitors or heat stress, making it a valuable model for studying cancer cell vulnerabilities linked to proteostasis imbalance. Moreover, given the HPV18-positive status and p53/Rb inactivation, this system allows exploration of how chaperone dysregulation intersects with viral oncoprotein-driven transformation and apoptosis evasion. The model also holds relevance for neurodegenerative disease research, where protein misfolding and aggregation are central pathological features.
Researchers can employ these polyclonal knockout cells in diverse experimental workflows, including immunoblotting to assess heat shock protein expression levels, immunofluorescence microscopy to visualize protein aggregates and subcellular localization of chaperones, and co-immunoprecipitation assays to probe interactions with Hsp70, Hsp90, or clathrin. Transcriptional responses mediated by HSF1 can be quantified via RT-qPCR, and cell viability assays under thermal or chemical stress provide functional readouts of chaperone capacity. This product is ideal for functional studies of chaperone biology, endocytosis, and cancer cell stress adaptation. For further technical details or assistance, please contact Ascent Research.
