ACSL4 Knockout T-47D Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of T-47D human breast cancer cells with targeted disruption of the ACSL4 gene. This bulk-knockout model provides a heterogeneous loss-of-function system that avoids clonal bias and is ideal for pooled functional screens and population-level assays. Generated via CRISPR/Cas9-mediated gene disruption, these polyclonal cells exhibit stable ablation of ACSL4 function, enabling robust investigation of ferroptosis and lipid metabolism pathways.
T-47D is a widely used ductal carcinoma cell line derived from a pleural effusion. It displays a luminal epithelial phenotype and expresses estrogen, progesterone, and androgen receptors, representing a triple-positive breast cancer model. This hormone-responsive background makes T-47D particularly suitable for studying endocrine-related tumor biology and how metabolic cell death pathways intersect with hormone signaling.
ACSL4 encodes a long-chain fatty acyl-CoA synthetase that preferentially activates arachidonic acid to arachidonoyl-CoA. This intermediate is subsequently utilized by LPCAT3 for incorporation into membrane phospholipids. Oxidation by lipoxygenases, notably ALOX15 and ALOX12, converts these phospholipids into lipid hydroperoxides that seed ferroptosis. The anti-ferroptotic enzyme GPX4 directly counters this process by reducing lipid hydroperoxides. ACSL4 is transcriptionally regulated by SP1, PPARG, SREBF1, NFE2L2, and TP53, and its expression responds to insulin and arachidonic acid levels. It also interacts with WWTR1/TAZ, integrating metabolic and oncogenic signals.
In T-47D cells, ACSL4 knockout prevents arachidonoyl-CoA formation, thereby blocking the lipid peroxidation cascade essential for ferroptosis execution. This confers resistance to ferroptosis inducers such as erastin and RSL3, providing a clean system to segregate ferroptotic cell death from apoptosis or necroptosis. The model is valuable for exploring how lipid metabolism and ferroptosis influence hormone therapy response in ER-positive breast cancer, aiding studies on drug resistance mechanisms.
Key applications include immunoblotting and RT-qPCR for expression validation, C11-BODIPY staining and flow cytometry to monitor lipid ROS, and viability assays with ferroptosis inducers. Functional assays such as colony formation, Transwell migration/invasion, xenograft models, and metabolomic profiling are also supported. The cells are suited for co-immunoprecipitation of ACSL4 interactors and phenotypic screening of ferroptosis-modulating compounds, facilitating biomarker discovery and anti-cancer drug development. For additional technical details, please reach out to our support team.
