The DNAJC16 Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the DNAJC16 gene in HeLa cells, creating a versatile loss-of-function model for investigating chaperone-mediated protein homeostasis. This product, offered as a polyclonal pool, contains a heterogeneous mixture of cells harboring CRISPR/Cas9-mediated target-gene disruption, enabling robust assessment of DNAJC16 function without the clonal selection biases inherent to monoclonal lines. The polyclonal format preserves population-level biological complexity and is particularly suitable for bulk biochemical, transcriptomic, and phenotypic assays where reproducible knockout effects are observed across the edited cell population.
HeLa cells, derived from an HPV18-positive cervical adenocarcinoma, are an established epithelial model widely employed in cancer biology, protein folding research, and stress response studies. Their transformed phenotype, high proliferative capacity, and well-characterized chaperone network render them an ideal host for examining the consequences of DNAJC16 deletion. The epithelial origin of HeLa cells also makes this model pertinent for interrogating how co-chaperone dysregulation impacts tissue-specific proteostasis and oncogenic signaling in carcinoma contexts.
DNAJC16 encodes a J-domain co-chaperone that selectively recruits HSP70 family proteins, principally HSPA8 (HSC70) and HSPA14, to client substrates, thereby facilitating protein folding, assembly, and intracellular trafficking. Acting downstream of the transcription factor HSF1 and cellular stress stimuli, DNAJC16 modulates chaperone cycles by stimulating HSP70 ATPase activity in conjunction with co-adaptors. Within the broader network, it functionally intersects with HSP90, the E3 ubiquitin ligase CHIP, and BAG family co-chaperones, which collectively determine client fate between folding, degradation, or aggregation. Knockout of DNAJC16 perturbs these interactions, potentially redirecting misfolded proteins toward aggregation or proteasomal clearance and altering signaling through downstream effectors such as apoptotic regulators and stress-activated kinases.
In the HeLa cellular context, DNAJC16 disruption is expected to compromise chaperone-mediated quality control, leading to the accumulation of unfolded protein clients and a heightened basal stress response. Given HeLa cells’ reliance on robust proteostasis to sustain rapid proliferation and survive oncogenic stress, this knockout model provides a tractable system for exploring how co-chaperone deficiency reshapes the proteotoxic landscape of cancer cells. It also offers insights into the role of J-domain proteins in the cellular stress response and their potential as therapeutic targets in malignancies characterized by protein folding vulnerabilities.
This product enables a wide range of research applications, including mechanistic studies of chaperone biology, proteostasis network analysis, and drug target validation. Researchers can employ co-immunoprecipitation to probe disrupted HSPA8-containing complexes, Western blotting to monitor changes in HSP70 client protein levels, and immunofluorescence to visualize protein aggregate formation. Functional assays such as flow cytometry for apoptosis induction and drug sensitivity profiling are readily applicable, while RNA-seq can identify downstream transcriptional changes in stress response genes. For further information, please contact Ascent Research.
