The Gpx4 Knockout PC-12 Cell Line is a CRISPR/Cas9-edited knockout cell line that disrupts the Gpx4 gene in the rat PC-12 cell line. This targeted disruption abolishes GPX4 protein expression, generating a loss-of-function model defective in lipid hydroperoxide detoxification. Consequently, cells are highly susceptible to ferroptosis induction by oxidative stress, making this line a robust platform for studying regulated necrotic cell death. The cell line is provided as a ready-to-use in vitro research tool and does not require additional editing or clonal isolation.
The PC-12 host cell line originates from a transplantable rat adrenal medullary pheochromocytoma and retains dual neuroendocrine and neuronal differentiation characteristics. After exposure to nerve growth factor (NGF), PC-12 cells cease proliferation, extend neurites, and acquire a sympathetic neuron-like phenotype. This established model is widely employed for studying neuronal differentiation, neurotrophin signaling, and neurosecretion, as well as oxidative stress-related neurodegeneration. The availability of the Gpx4 knockout in this background allows investigators to examine ferroptotic processes in both undifferentiated and neuronally differentiated cells.
GPX4 (glutathione peroxidase 4) is a selenocysteine-containing enzyme that catalyzes the reduction of complex lipid hydroperoxides to lipid alcohols, using glutathione (GSH) as an electron donor. This reaction is the principal cellular mechanism for blocking ferroptosis??an iron-dependent form of regulated necrosis driven by lipid peroxidation. GPX4 expression is transcriptionally regulated by NFE2L2 (NRF2), ATF4, and TP53, and its activity is selenium-dependent. In the ferroptosis pathway, GPX4 functions downstream of the cystine/glutamate transporter SLC7A11, which supplies cysteine for GSH synthesis, and lipid remodeling enzymes ACSL4 and LPCAT3 that generate peroxidation substrates. Thus, GPX4 sits at a critical node integrating glutathione metabolism, iron homeostasis, and lipid oxidation.
In the Gpx4 knockout PC-12 line, the absence of GPX4 renders cells unable to neutralize lipid peroxides, culminating in rapid ferroptotic death upon exposure to ferroptosis inducers such as erastin or RSL3. Both undifferentiated and NGF-differentiated knockout cells accumulate lipid ROS and exhibit hallmarks of ferroptosis, providing a neural model to dissect cell death signaling. This system is particularly relevant for investigating ferroptosis contribution to cerebral ischemia-reperfusion injury, age-related neurodegeneration, and neurotoxicant exposure, where neuronal oxidative damage is implicated.
Applications include high-throughput screening of ferroptosis modulators, lipid peroxidation measurement with C11-BODIPY, and dose-response assays with erastin, RSL3, or other ferroptosis triggers. Standard biochemical readouts such as intracellular GSH quantification, ROS detection (H2DCFDA, MitoSOX), and ferroptosis inhibitor rescue (liproxstatin-1, deferoxamine) are fully supported. Western blotting for GPX4, ACSL4, and SLC7A11, as well as RT-qPCR analysis of ferroptosis marker genes, can be performed. For additional technical information, please contact Ascent Research.
