The ACSL4 Knockout KYSE-150 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the ACSL4 gene in the KYSE-150 human esophageal squamous cell carcinoma line. This product offers researchers a versatile loss-of-function model that eliminates ACSL4 expression across a mixed genetic background, avoiding clonal artifacts while enabling consistent pathway interrogation. It is designed for investigations into ferroptosis regulation, lipid metabolism, and oncogenic signaling.
KYSE-150 cells are derived from a poorly differentiated esophageal squamous cell carcinoma and carry a TP53 mutation, conferring a highly invasive phenotype. This well-established cell line models aggressive tumor behavior, including metastatic propensity and therapeutic resistance. The compromised p53 status is particularly relevant as p53 transcriptionally represses ACSL4; thus, the knockout system permits dissection of p53-dependent and independent regulation of ferroptosis in a clinically relevant context.
The ACSL4 enzyme catalyzes the conversion of long-chain polyunsaturated fatty acids??especially arachidonic acid??to their acyl-CoA derivatives, a prerequisite for their incorporation into membrane phospholipids via LPCAT3. This activity enriches cellular membranes with peroxidation-prone acyl chains, which, upon oxidative challenge, are oxygenated by lipoxygenases such as ALOX5 and ALOX12 to generate phospholipid hydroperoxides that execute ferroptosis. ACSL4 expression is controlled by upstream regulators including p53 (represses), SREBP1c, ATF4, and PPAR??, and its function opposes the antioxidant defense mediated by SLC7A11 and GPX4. By producing arachidonoyl-CoA and promoting phospholipid remodeling, ACSL4 acts as a sentinel for ferroptotic sensitivity.
Knocking out ACSL4 in KYSE-150 cells is expected to abrogate ferroptotic cell death triggered by erastin or RSL3, making this model instrumental for studying ferroptosis resistance mechanisms. The aggressive, TP53-mutant background allows researchers to examine how loss of this pro-ferroptotic factor alters lipid metabolism, tumor invasiveness, and drug sensitivity. This cellular tool enables discrimination between ferroptosis and other cell death modalities, elucidating ACSL4-specific contributions to cancer pathophysiology.
These polyclonal knockout cells are ideal for ferroptosis-focused assays including C11-BODIPY lipid peroxidation staining, viability testing with erastin/RSL3, and western blotting for ACSL4 and GPX4. LC-MS lipidomics, glutathione and iron measurement, and RT-qPCR for ferroptosis markers complement the workflow. The model facilitates high-throughput screening of ferroptosis modulators and preclinical evaluation of lipid metabolism-targeted therapies in esophageal cancer. For technical specifications or to order, please contact Ascent Research.
