The GSS Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the glutathione synthetase (GSS) gene in the human non-small cell lung cancer (NSCLC) cell line NCI-H1975. This product provides a heterogeneous pool of cells with GSS gene disruption, enabling loss-of-function studies without clonal selection. The polyclonal format maintains population diversity while abrogating GSS expression, suitable for redox biology and tumor vulnerability studies in a lung adenocarcinoma background.
The NCI-H1975 cell line originates from the pleural effusion of a nonsmoking female with lung adenocarcinoma and is a widely used NSCLC model. It harbors EGFR L858R and T790M mutations, conferring constitutive kinase activity and resistance to first-generation tyrosine kinase inhibitors, respectively. These genetic features make the cell line particularly relevant for studying signaling dependencies, metabolic adaptation, and drug resistance in EGFR-mutant lung cancer.
GSS encodes glutathione synthetase, catalyzing the final step of glutathione (GSH) synthesis by ligating gamma-glutamylcysteine and glycine. GSH is a critical antioxidant that serves as a cofactor for GPX4 to reduce lipid peroxides and prevent ferroptosis, and for glutathione S-transferases in detoxification. GSS is transcriptionally regulated by NFE2L2 (Nrf2) under oxidative stress, and functions downstream of GCL and upstream of glutathione reductase. The pathway integrates with SLC7A11-mediated cystine import, positioning GSS at a central node in ferroptosis defense and cellular redox homeostasis.
In NCI-H1975 cells, GSS knockout depletes GSH, impairing redox balance and sensitizing cells to oxidative stress and ferroptosis. EGFR-mutant NSCLC cells often rely on heightened antioxidant capacity to manage oncogenic stress, making the GSH system a therapeutically relevant vulnerability. This model enables dissection of redox-dependent cell death mechanisms and evaluation of ferroptosis-targeting strategies in a relevant lung adenocarcinoma background.
The polyclonal knockout cells are suitable for functional assays including GSH/GSSG ratio measurements, lipid peroxidation detection by C11-BODIPY, and ROS quantification by flow cytometry. Cell viability responses to ferroptosis inducers (erastin, RSL3) can be assessed in colony formation and xenograft models. Western blotting for GPX4 and SLC7A11 confirms pathway involvement. For further information, please contact Ascent Research.
