The ACSL4 Knockout DLD-1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout population derived from the DLD-1 human colorectal adenocarcinoma cell line. These cells carry targeted ACSL4 gene disruption generated via CRISPR/Cas9-mediated genome editing, providing a loss-of-function model to investigate ferroptosis and lipid metabolism. The polyclonal format encompasses a heterogeneous mixture of edited alleles, minimizing clonal selection bias and enabling studies of ACSL4 function in a genetically diverse cellular context.
DLD-1 is a well-established epithelial colorectal adenocarcinoma cell line isolated from a male patient. It serves as a robust model for colorectal cancer biology, including tumorigenesis, metastasis, and oncogenic signaling. DLD-1 cells harbor characteristic mutations that drive malignant phenotypes, making them an ideal host for evaluating the impact of gene disruption on cancer-related processes. Their reproducible growth and extensive characterization support rigorous functional experimentation.
ACSL4 encodes an acyl-CoA synthetase that activates long-chain polyunsaturated fatty acids (PUFAs), converting them to acyl-CoA esters. This step facilitates the incorporation of PUFAs into membrane phospholipids by LPCAT3, generating phosphatidylethanolamine-PUFA (PE-PUFA) species that prime membranes for lipid peroxidation??a critical event in ferroptotic cell death. ACSL4 is transcriptionally regulated by SREBP1 and PPAR??, and its activity is influenced by insulin, growth factors, and lipid availability. ACSL4 operates upstream of lipid peroxide accumulation and functionally interacts with GPX4, LPCAT3, and lipoxygenases (LOXs). In the ferroptosis pathway, ACSL4-dependent lipid remodeling increases sensitivity to oxidative damage, which is counterbalanced by GPX4-mediated reduction of lipid hydroperoxides.
Within the DLD-1 colorectal cancer context, ACSL4 knockout offers a system to decipher ferroptosis susceptibility and tumor cell vulnerabilities. Colorectal tumors often exhibit dysregulated lipid metabolism and redox homeostasis; thus, this model allows dissection of how ACSL4-driven lipid peroxidation affects cancer cell survival, metastatic capacity, and response to ferroptosis-inducing chemotherapeutics. The polyclonal knockout configuration captures population-level heterogeneity, reflecting the mosaic nature of tumor gene editing and enabling robust assessment of cell death pathway engagement.
This product supports a wide array of research applications, including ferroptosis mechanism exploration, lipid metabolism profiling, and anticancer drug sensitivity screening. Key assays include western blotting for ACSL4 protein loss, RT-qPCR for transcript confirmation, C11-BODIPY staining to quantify lipid peroxidation, and cell viability measurements following ferroptosis induction with erastin or RSL3. Immunofluorescence can localize lipid peroxidation events or protein distribution. For additional technical details or support, please contact Ascent Research.
