The ACSL4 Knockout AGS Polyclonal Cells are a genetically engineered human cell population created by CRISPR/Cas9-mediated disruption of the ACSL4 gene within the AGS gastric adenocarcinoma epithelial cell line. This polyclonal knockout product offers a pooled loss-of-function model for studying acyl-CoA synthetase long-chain family member 4 (ACSL4) functions in lipid metabolism and ferroptosis, without isolation of a single clone.
AGS cells were derived from a human gastric adenocarcinoma and retain key epithelial characteristics, including adhesion and polarization, commonly utilized in gastric cancer pathogenesis and drug response studies. Their origin makes them a relevant model for investigating molecular events in gastric mucosal biology and carcinogenesis.
ACSL4 catalyzes the ligation of long-chain polyunsaturated fatty acids (PUFAs), notably arachidonic acid, with coenzyme A (CoA), enabling their esterification into phosphatidylethanolamine (PE) by lysophosphatidylcholine acyltransferase 3 (LPCAT3). The resulting PUFA-PE species are susceptible to iron-dependent peroxidation by lipoxygenases (ALOXs), generating lipid peroxides that drive ferroptotic cell death. GPX4 reduces these peroxides, thereby serving as a key inhibitor of ferroptosis. Upstream, ACSL4 expression is transcriptionally regulated by sterol regulatory element-binding protein 2 (SREBP2) and the tumor suppressor TP53, and can be induced by lipopolysaccharide (LPS) and oxidative stress. Downstream execution involves accumulation of lipid peroxides, glutathione (GSH) depletion, and GPX4 inhibition, with ACSL4 interacting directly with arachidonic acid, CoA, and the ferroptosis regulators GPX4 and iron.
In gastric adenocarcinoma, lipid metabolic reprogramming and ferroptosis sensitivity are clinically relevant, as ACSL4 expression levels can influence tumor cell fate under oxidative stress and chemotherapeutic exposure. The AGS polyclonal knockout model provides a physiologically pertinent system to dissect ACSL4-dependent lipid remodeling and its impact on ferroptosis sensitivity, revealing potential vulnerabilities for therapeutic intervention in gastric and other ACSL4-expressing cancers.
This ACSL4 knockout polyclonal cell pool is suitable for ferroptosis research through assays such as C11-BODIPY lipid peroxidation staining, viability testing with ferroptosis inducers (e.g., erastin, RSL3), and immunoblotting for ACSL4 and GPX4. Transcript analysis by RT-qPCR, iron content measurement, GSH quantification, and mitochondrial ultrastructural examination by transmission electron microscopy further expand experimental scope. The model supports identification of ferroptosis-regulating compounds, validation of lipid metabolic biomarkers, and elucidation of ACSL4-mediated signaling in gastric cancer. For further details, please contact Ascent Research.
