The GPX8 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population generated from human embryonic kidney HEK293T cells via disruption of the endogenous GPX8 gene. This heterogeneous pool contains diverse editing events, providing a robust model for loss-of-function studies without clonal selection bias. The cells are supplied as an expansion-ready culture, suitable for immediate application in biochemical, genetic, or pharmacological assays investigating GPX8-dependent processes.
The parental HEK293T line originates from human embryonic kidney epithelium and is stably transformed with the SV40 large T antigen, conferring rapid growth and superior transfection efficiency. This cell line is a staple for heterologous expression, viral production, and CRISPR-based genome engineering. HEK293T cells possess an intact glutathione antioxidant system and are responsive to ferroptotic stimuli, making them a suitable platform for investigating redox-sensitive proteins such as GPX8 in a well-characterized human kidney context.
GPX8 is an endoplasmic reticulum-localized glutathione peroxidase that catalyzes the reduction of hydrogen peroxide and organic hydroperoxides at the expense of reduced glutathione (GSH). Its expression is regulated by NFE2L2/Nrf2 in response to oxidative stress and by ATF4 during ER stress, and its activity requires selenium. GPX8 cooperates with glutathione reductase (GR), which recycles GSH from oxidized glutathione using NADPH, and functionally intersects with thioredoxin and peroxiredoxin networks. By detoxifying lipid hydroperoxides, GPX8 suppresses ferroptosis, and its disruption leads to elevated ROS and lipid peroxidation.
The HEK293T background, with its robust glutathione metabolism and active lipid routes, is well-suited to reveal the consequences of GPX8 loss on endoplasmic reticulum redox homeostasis and ferroptosis sensitivity. The polyclonal knockout population enables assessment of global antioxidant capacity and identifies adaptive pathway alterations that might be masked in monoclonal isolates. This model assists in evaluating GPX8??s protective roles against oxidative chemotherapeutics and its contribution to ferroptosis resistance in cancer cells, as well as in modeling renal epithelial vulnerability to ischemia-reperfusion and nephrotoxic insults.
These knockout cells are extensively used to dissect GPX8-mediated ferroptosis suppression through lipid peroxidation assays (C11-BODIPY) and ROS detection (DCFDA). They facilitate dose-response studies with ferroptosis inducers like erastin and enable glutathione quantification to probe redox dynamics. Screening for GPX8-specific antioxidants or synthetic lethal partners is streamlined by the cell population??s heterogeneity. Standard validation includes Western blotting and RT-qPCR. For further technical information or customized orders, please contact Ascent Research.
