CCS Knockout HeLa Polyclonal Cells constitute a CRISPR/Cas9-mediated polyclonal knockout cell population designed for the disruption of the CCS gene in a HeLa background. This product is supplied as a heterogeneous pool of edited cells, enabling functional studies of CCS-dependent copper homeostasis without clonal selection. The use of a polyclonal knockout format helps mitigate clonal artifacts and provides a population-level representation of gene disruption effects. These cells serve as a versatile loss-of-function model for investigating the role of the copper chaperone CCS in antioxidant defense and copper delivery to superoxide dismutase 1 (SOD1).
The host HeLa cell line is a human cervical adenocarcinoma line that is HPV18-positive and immortalized, widely employed in biomedical research due to its robust growth, epithelial morphology, and cancerous origin. HeLa cells exhibit dysregulated redox signaling and serve as a well-characterized platform for examining oxidative stress responses, metal homeostasis, and oncogenic pathways. Their established use in cell biology, biochemistry, and drug discovery makes them an ideal host for CRISPR-based knockout models.
CCS (copper chaperone for superoxide dismutase) is a metallochaperone that specifically binds and delivers copper ions to SOD1, a critical event for SOD1 enzymatic activation. SOD1 subsequently catalyzes the dismutation of superoxide radicals into oxygen and hydrogen peroxide. The CCS gene is transcriptionally upregulated by the oxidative stress-responsive transcription factor NFE2L2 (Nrf2), linking it to cellular antioxidant programs. CCS directly interacts with SOD1 and copper ions, and its function is essential for superoxide detoxification. Disruption of CCS therefore results in inactive SOD1, accumulation of superoxide, and heightened oxidative stress.
In the HeLa cellular context, CCS knockout provides a powerful tool to dissect the interplay between copper metabolism and redox homeostasis in a cancer-relevant setting. Because cancer cells often exhibit elevated basal reactive oxygen species and altered metal ion trafficking, the loss of CCS can reveal vulnerabilities associated with SOD1 dysfunction and copper dependency. This model is pertinent for research into amyotrophic lateral sclerosis (ALS), as CCS-SOD1 pathway perturbations are central to familial ALS pathology, but it also extends to studies of copper metabolism disorders and general oxidative stress biology.
This knockout product is suited for a variety of experimental applications, including western blotting to confirm CCS protein loss, SOD1 activity assays to assess copper loading defects, reactive oxygen species detection using DCFDA, cell viability measurements under oxidative challenge, immunofluorescence for CCS subcellular localization, RT-qPCR for transcriptional analysis, and copper uptake assays. By providing a polyclonal knockout population, researchers can examine these endpoints in a genetically diverse background that mimics physiological heterogeneity. For further information or custom requests, please contact Ascent Research.
