The GPX4 Knockout AGS Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human gastric adenocarcinoma cell line AGS, featuring targeted disruption of the GPX4 gene. This loss-of-function model provides a robust system for investigating GPX4-dependent mechanisms in ferroptosis and oxidative stress response, leveraging the genetic heterogeneity of a polyclonal population while maintaining stable gene ablation. The polyclonal format enables population-level studies without the constraints of single-cell clones, suitable for broad experimental applications in cancer biology and redox biochemistry.
AGS is a well-characterized epithelial cell line established from a primary human gastric adenocarcinoma, widely employed as an in vitro model for gastric cancer. It retains key molecular features of gastric carcinogenesis, including dysregulated proliferation, altered signal transduction, and aberrant oxidative stress handling. The gastric adenocarcinoma background is particularly relevant for ferroptosis studies, as gastric tumors frequently exhibit imbalanced redox homeostasis and lipid metabolism, rendering the AGS line an appropriate host to assess GPX4 function in tumor cell survival and drug sensitivity.
GPX4 encodes a glutathione peroxidase that catalyzes the reduction of phospholipid hydroperoxides to lipid alcohols using glutathione (GSH) as an essential cofactor, thereby suppressing iron-dependent lipid peroxidation and blocking ferroptotic cell death. Expression of GPX4 is transcriptionally regulated by the oxidative stress sensor NRF2 (NFE2L2), and its enzymatic activity depends on selenium incorporation, cysteine availability for GSH synthesis, and sufficient intracellular glutathione levels. GPX4 directly interacts with GSH and lipid substrates, functioning downstream of the cystine/glutamate antiporter SLC7A11, which imports cystine for glutathione biosynthesis. The enzyme acts in opposition to pro-ferroptotic factors such as ACSL4 and ALOX15, which promote the generation of phospholipid hydroperoxides. Disruption of GPX4 in the AGS knockout cells eliminates this protective axis, rendering cells acutely sensitive to ferroptosis induction and consequent lipid oxidative damage.
Ablation of GPX4 in AGS gastric adenocarcinoma cells creates an indispensable model for dissecting ferroptosis regulation within a clinically relevant gastric cancer context. The knockout sensitizes cells to ferroptotic death upon lipid peroxide accumulation, enabling direct interrogation of the protective role of GPX4 against oxidative injury and its contribution to mitochondrial integrity maintenance. This system is instrumental for evaluating how GPX4 loss influences tumor cell viability under glutathione-depleting conditions, for screening ferroptosis-inducing agents such as RSL3 or erastin, and for studying adaptive resistance mechanisms. Moreover, the model extends to investigations of ferroptosis-related pathologies, including neurodegenerative disorders where lipid peroxidation is a key pathogenic driver.
Typical applications of GPX4 Knockout AGS Polyclonal Cells encompass ferroptosis induction assays using standardized inducers coupled with cell viability quantitation to measure death sensitivity. Lipid peroxidation levels are readily assessed via C11-BODIPY staining and flow cytometry, while glutathione content is monitored by colorimetric or fluorometric methods. Western blotting and RT-qPCR confirm GPX4 ablation and probe compensatory pathways, including NRF2 target gene expression. Additional uses include drug combination screens, genetic rescue experiments, and transcriptomic profiling of ferroptosis-responsive networks. For further technical details and ordering information, please contact Ascent Research.
