The DNAJC7 Knockout HeLa Polyclonal Cells product provides a robust and versatile tool for studying the co-chaperone DNAJC7 in cellular stress and protein homeostasis. This CRISPR/Cas9-edited polyclonal knockout population, targeting the DNAJC7 gene in HeLa cells, is generated through CRISPR/Cas9-mediated gene disruption, yielding a heterogeneous loss-of-function model without the need for single-cell cloning. It is designed for pooled screening and bulk assays, retaining genetic diversity while achieving effective target-gene knockout in a widely used human cervical cancer cell line.
HeLa cells are an immortalized human cervical epithelial carcinoma line, established from a patient with adenocarcinoma and containing integrated HPV18 sequences. They are extensively employed in oncology, virology, and cell signaling research due to their rapid proliferation and responsiveness to genetic manipulation. Their HPV oncoproteins (E6 and E7) disrupt p53 and Rb tumor suppressor pathways, yet major stress signaling and chaperone networks remain functional, making them a relevant host for investigating DNAJC7-mediated protein quality control.
DNAJC7 is a TPR-domain co-chaperone that directly binds to the C-terminal tails of HSPA8/HSC70 and HSP90AA1, facilitating client protein folding, assembly, and degradation. It recruits the E3 ubiquitin ligase STUB1/CHIP to ubiquitinate terminally misfolded clients, thereby linking chaperone activity to the ubiquitin-proteasome system. DNAJC7 expression is upregulated by heat shock via HSF1 and by endoplasmic reticulum stress through the unfolded protein response sensors ATF6, IRE1, and PERK. Its activity modulates the conformation and stability of Hsp70/Hsp90 client proteins, including kinases and steroid receptors, and influences apoptosis by regulating BAX, BCL2, and caspase activation. Additionally, DNAJC7 interacts with PARK2/Parkin, implicating it in mitochondrial protein quality control and mitophagy.
In HeLa cervical cancer cells, DNAJC7 knockout disrupts cellular proteostasis, leading to accumulation of aggregation-prone proteins and sensitizing cells to apoptosis under stress conditions. Cancer cells rely heavily on chaperone networks to handle oncogenic proteotoxic stress; thus, loss of DNAJC7 may impair survival and enhance sensitivity to chemotherapeutic agents. Moreover, the DNAJC7 gene has been identified as a genetic risk factor for amyotrophic lateral sclerosis (ALS), so this knockout population also serves as a tool to investigate mechanisms of protein aggregation and neurodegeneration. The model enables dissection of the balance between chaperone-mediated refolding and degradation pathways.
This polyclonal knockout population supports a wide array of applications, including flow cytometry for apoptosis, western blotting for Hsp70/Hsp90 targets, RT-qPCR for chaperone gene expression, cell viability assays, proteasome activity measurements, co-immunoprecipitation for protein interactions, and immunofluorescence for visualizing protein aggregation. Researchers can use this tool to screen for small-molecule modulators of the Hsp70/Hsp90 system, evaluate client protein stability, and investigate stress response mechanisms under conditions such as heat shock or ER stress. For further information and technical support, please contact Ascent Research.
