ACSL4 Knockout 769-P Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the 769-P human clear cell renal cell carcinoma (ccRCC) line, featuring a targeted disruption of the ACSL4 gene. This loss-of-function model enables investigation of ACSL4-dependent processes without detailed genetic characterization, as the polyclonal format maintains the allelic diversity of pooled gene disruption. The cells provide a reliable platform for studying ferroptosis and lipid metabolism in a cancer-relevant epithelial context.
The parental 769-P line, isolated from a primary ccRCC tumor, exhibits epithelial morphology and retains metabolic hallmarks of kidney cancer, including dysregulated fatty acid metabolism. Widely used in renal cancer research, these cells respond to ferroptosis inducers and are suitable for modeling ACSL4??s role in oncogenic lipid handling and cell death pathways.
ACSL4 catalyzes the ATP-dependent activation of long-chain polyunsaturated fatty acids (PUFAs) to acyl-CoA esters, a prerequisite for their incorporation into membrane phospholipids by LPCAT3. This enrichment of PUFA-phosphatidylethanolamines renders membranes susceptible to peroxidation by lipoxygenases (e.g., ALOX15) in the presence of labile iron (Fe2+), generating lipid hydroperoxides that execute ferroptosis. ACSL4 expression is positively regulated by SREBP1, PPAR??, TFAP2C, and hypoxia, and it functions upstream of ferroptosis execution by promoting pro-ferroptotic lipid synthesis. The enzyme is functionally opposed by GPX4 and the system xc? (SLC7A11)-glutathione axis, and collaborates with interacting partners like ACSL1 and LPCAT3 in lipid metabolic networks.
In 769-P ccRCC cells, ACSL4 disruption creates a powerful model to explore ferroptosis vulnerability and lipid metabolic reprogramming. Given that ccRCC frequently exhibits ferroptosis resistance, this knockout allows direct assessment of how loss of PUFA activation alters sensitivity to erastin, RSL3, and other ferroptosis-inducing agents. It also enables study of compensatory pathways involving ACSL1 or LPCAT3 and the impact on membrane lipid composition.
These cells support diverse assays: ferroptosis induction with erastin/RSL3 coupled to viability measurements (MTT) and lipid peroxidation detection (C11-BODIPY); western blotting for ACSL4, GPX4; RT-qPCR; iron quantification; and ROS detection. They are applicable in drug sensitivity profiling, ischemia-reperfusion injury models, and neurodegeneration studies. For more details, contact Ascent Research.
