ACSL4 Knockout 143B Polyclonal Cells are a CRISPR/Cas9-mediated polyclonal knockout population derived from the 143B human osteosarcoma cell line. The product delivers a heterogeneous cell pool with disrupted ACSL4 expression, obtained by transient introduction of CRISPR components followed by selection. This polyclonal format avoids the limitations of single-cell cloning and captures the functional diversity inherent in a gene-edited population, providing a robust platform for studying ACSL4??s role in lipid metabolism and programmed cell death.
The 143B cell line is a TP53-mutant osteosarcoma model characterized by aggressive growth, anchorage independence, and tumorigenicity in xenografts. Widely used in cancer biology, these cells offer a clinically relevant system for examining mechanisms of tumor survival, metastasis, and therapy resistance. Their osteosarcoma origin makes them particularly suited for investigating how alterations in lipid metabolism and oxidative stress pathways influence sarcoma progression.
ACSL4 encodes a long-chain acyl-CoA synthetase that preferentially activates arachidonic acid to arachidonoyl-CoA, driving the esterification of polyunsaturated fatty acids into membrane phospholipids. This enzymatic step is essential for generating ferroptosis-sensitive lipid species. ACSL4 expression is regulated by SREBF1, TFAP2C, and SP1, and its product serves as substrate for LPCAT3-mediated phospholipid remodeling. Downstream, arachidonoyl-containing phosphatidylethanolamines are oxidized by ALOX15 and other lipoxygenases, promoting lipid peroxide accumulation that is counteracted by GPX4 and the System Xc- cystine importer.
In 143B osteosarcoma cells, ACSL4 knockout abrogates the production of oxidation-prone phospholipids, thereby conferring resistance to ferroptosis inducers such as erastin and RSL3. This model permits dissection of the interplay between p53 status and ferroptosis sensitivity, as TP53 mutations may influence ACSL4 expression and lipid metabolic rewiring. The polyclonal knockout population enables assessment of heterogeneous ferroptotic responses within a cancer cell pool, a phenomenon that may impact therapeutic targeting strategies.
Researchers can employ this product in lipid peroxidation assays using C11-BODIPY, cell viability assessments with ferroptosis modulators, and molecular analyses via western blotting for ACSL4, GPX4, and related proteins. Functional assays such as Transwell migration/invasion, colony formation, and xenograft tumor models extend the utility to metastasis and in vivo therapeutic studies. These applications facilitate ferroptosis research, osteosarcoma drug testing, and investigation of ischemia-reperfusion injury mechanisms. For technical inquiries or order details, please contact Ascent Research.
