The GSS Knockout SK-HEP-1 Polyclonal Cells are a heterogeneous pool of SK-HEP-1 cells with CRISPR/Cas9-mediated gene disruption at the GSS locus. This polyclonal knockout model eliminates glutathione synthetase expression, providing a loss-of-function system for studying glutathione metabolism and redox homeostasis. By avoiding clonal isolation, the product maintains population-level diversity, suitable for pooled screening and robust phenotypic analyses.
SK-HEP-1 is a human epithelial hepatocellular carcinoma cell line derived from ascitic fluid of a liver adenocarcinoma patient. It is extensively used in hepatic cancer research due to its aggressive malignant characteristics and metabolic dysregulation. The line’s hepatic origin and mesenchymal features make it relevant for investigating redox adaptation and ferroptosis in liver tumors.
GSS encodes glutathione synthetase, which catalyzes the ATP-dependent ligation of gamma-glutamylcysteine and glycine to form glutathione (GSH). This step is crucial for maintaining intracellular GSH, a key antioxidant and cofactor. GSS is regulated transcriptionally by NRF2 and ATF4 during stress responses. Downstream, GSH serves as a substrate for GPX4 to reduce lipid hydroperoxides, thereby inhibiting ferroptosis. GSH also supports glutathione S-transferases and glutaredoxins. The biosynthetic pathway involves GCLC and GCLM, making GSS essential for cellular redox control.
Hepatocellular carcinoma cells frequently upregulate glutathione synthesis to counteract oxidative stress and chemotherapeutics. Disruption of GSS in SK-HEP-1 cells abrogates GSH production, sensitizing them to ferroptosis induction and oxidative damage. This model enables dissection of GSS-dependent drug resistance mechanisms and evaluation of alternative redox pathways, such as cystine import, in liver cancer biology.
This knockout product is applicable for glutathione quantification, ROS flow cytometry, western blotting of GSS and GPX4, and lipid peroxidation assays using C11-BODIPY. Ferroptosis can be induced with erastin, and cell viability measured under oxidative challenge. The cells also support studies on NRF2/ATF4 signaling. For further information, please contact Ascent Research.
