The ACSL4 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from HT29 colorectal adenocarcinoma cells, featuring targeted disruption of the ACSL4 gene. This loss-of-function model eliminates ACSL4 protein expression, enabling ferroptosis and lipid metabolism studies. The polyclonal nature captures diverse editing events, providing a robust system for gene function analysis without single-cell cloning.
The HT29 cell line is a human colorectal adenocarcinoma model with epithelial morphology, originating from a primary tumor. It is widely used for intestinal epithelial and colorectal cancer research, retaining key signaling pathways like Wnt/??-catenin and lipid metabolism networks. HT29 cells are suitable for studying drug responses, tumor behavior, and antioxidant defenses, providing an optimal context for ACSL4 knockout studies.
ACSL4 catalyzes the ligation of long-chain PUFAs like arachidonic acid to CoA, enabling their incorporation into membrane phospholipids. This activity is regulated by SREBP1/2, PPAR??/??, LXR, and Wnt/??-catenin, promoting phospholipid PUFA enrichment. ACSL4 drives ferroptosis by sensitizing membranes to ALOX5/ALOX15-mediated peroxidation, generating lipid hydroperoxides. When GPX4 activity is impaired, ACSL4-dependent lipid peroxidation causes ferroptotic cell death. Additionally, ACSL4 contributes to eicosanoid synthesis and interacts with GPX4 and ACSL1 within the glutathione-dependent peroxide scavenging system, underscoring its role in lipid metabolism and regulated necrosis.
In HT29 colorectal cancer cells, ACSL4 knockout provides a tool to dissect ferroptosis susceptibility and lipid reprogramming. Colorectal cancers show altered lipid profiles and variable ferroptosis sensitivity; ACSL4 expression is linked to therapeutic resistance. Removing ACSL4 allows investigation of PUFA metabolism effects on tumor growth, metastasis, and drug response. This model also enables study of oncogenic signaling (Wnt/??-catenin) crosstalk with lipid peroxidation pathways, revealing potential vulnerabilities.
Researchers can use these cells for ferroptosis induction assays (e.g., erastin/RSL3), lipid peroxidation C11-BODIPY staining, cell viability tests, and phospholipidomic profiling. They support western blotting and RT-qPCR to verify pathway alterations, plus combinatorial studies on drug resistance or antioxidant defense. This model helps explore ACSL4??s role in colorectal cancer and its therapeutic potential. For further information, please contact Ascent Research.
