This product comprises a CRISPR/Cas9-edited polyclonal knockout cell population of Homo sapiens Ca Ski cells, in which the GPX4 gene has been disrupted to establish a loss-of-function model. The polyclonal pool contains a heterogeneous mixture of edits, enabling the study of GPX4-dependent functions without clonal selection artifacts. This format is suitable for investigating gene-dosage effects and population-level responses in a cervical carcinoma background, providing a physiologically relevant system for ferroptosis and oxidative stress research.
Ca Ski is a well-characterized human cervical epithelial carcinoma cell line derived from a metastatic site in the small intestine. These adherent cells harbor integrated HPV16 DNA and express the viral oncoproteins E6 and E7, which inactivate p53 and retinoblastoma protein, respectively, driving malignant transformation. The line serves as a widely used model for HPV16-associated cervical carcinogenesis, exhibiting molecular features of squamous cell carcinoma and intact glutathione metabolism, making it an appropriate host for dissecting GPX4??s role in redox balance and cell survival.
GPX4 (glutathione peroxidase 4) is a phospholipid hydroperoxidase that catalyzes the glutathione-dependent reduction of lipid hydroperoxides, particularly phosphatidylethanolamine hydroperoxides (PUFA-PE-OOH), into non-toxic lipid alcohols, thereby preventing iron-dependent oxidative membrane damage and inhibiting ferroptotic cell death. Its activity is regulated by upstream signals including NRF2-mediated transcription, selenium bioavailability, and mTORC1, while p53 can transcriptionally repress SLC7A11, the substrate-specific subunit of the cystine/glutamate antiporter System Xc?C, modulating glutathione synthesis. GPX4 function is intimately linked to ACSL4 and ALOX family members that promote lipid hydroperoxide generation, and it interacts with SECISBP2 for selenocysteine incorporation and SBP1 for activity. The enzyme thus sits at the nexus of a pathway involving SLC7A11/SLC3A2-mediated cystine import, glutathione biosynthesis, and lipid peroxide detoxification.
In the Ca Ski background, disruption of GPX4 is expected to sensitize cells to ferroptosis inducers such as erastin (System Xc?C inhibitor) and RSL3 (direct GPX4 inhibitor), revealing the reliance of HPV-positive cervical carcinoma cells on this axis for survival under oxidative stress. Since HPV oncoproteins can alter redox homeostasis and promote lipid metabolism reprogramming, this knockout population provides a platform to examine how viral transformation rewires ferroptosis susceptibility. It also enables elucidation of compensatory mechanisms that may arise in a polyclonal setting, such as upregulation of alternative antioxidant pathways.
This GPX4 knockout Ca Ski polyclonal cell model is a versatile tool for ferroptosis-centered research. Applications include mechanistic studies of oxidative cell death pathways, high-throughput screening of ferroptosis-modulating compounds, and evaluation of chemotherapeutic synergy in HPV-driven cancers. End users can assess protein expression via Western blotting for GPX4 and the lipid peroxidation marker 4-HNE, quantify lipid peroxidation using the C11-BODIPY probe by flow cytometry, and measure cell viability after treatment with erastin or RSL3. Glutathione levels can be monitored, and transcriptional profiling by RT-qPCR (GPX4, SLC7A11) or RNA-seq can uncover downstream molecular changes. For further details on this product, please contact Ascent Research.
