The ACSL5 Knockout HT29 Polyclonal Cells consist of a heterogeneous population of HT29 human colorectal adenocarcinoma cells carrying CRISPR/Cas9-mediated disruption of the ACSL5 gene. This polyclonal knockout format preserves genetic variability and avoids clonal selection biases, offering a robust model for functional studies of ACSL5 in a relevant epithelial tumor context. The knockout population enables loss-of-function analysis in a setting that more closely mimics the heterogeneity of tumor cell populations.
HT29 cells are a widely used human colorectal adenocarcinoma model originating from a 44-year-old female. These adherent epithelial cells are KRAS wild-type and harbor a TP53 mutation. Importantly, they can differentiate into enterocyte-like cells under glucose deprivation, providing a unique system for studying intestinal epithelial differentiation and the metabolic adaptations of colorectal cancer cells.
ACSL5 encodes a long-chain acyl-CoA synthetase that activates long-chain fatty acids to acyl-CoA, a critical step for their utilization in triglyceride synthesis, phospholipid production, and ??-oxidation. Its expression is regulated by PPAR??, PPAR??, SREBP1, LXR, and insulin, and it functionally interacts with FABP1, CPT1A, DGAT1, and ACSL4 to channel fatty acids into storage or oxidation. In the acyl-CoA pathway, FATP4 imports fatty acids which are activated by ACSL5, then processed by GPAT, AGPAT, and DGAT for lipid droplet formation, or by CPT1A and ACOX1 for ??-oxidation, influencing PPAR?? signaling.
Disruption of ACSL5 in HT29 cells is anticipated to reduce acyl-CoA ester production, thereby limiting lipid droplet formation and altering phospholipid distributions. The TP53 mutant and glucose deprivation-responsive differentiation capacity of these cells make the knockout model particularly valuable for studying the contribution of ACSL5-driven lipid metabolism to tumor cell proliferation, survival, and enterocyte-like differentiation. This system provides a platform to examine metabolic vulnerabilities in colorectal cancer and to assess the impact on tumor progression.
This knockout model is suitable for quantifying fatty acid uptake, triglyceride accumulation, and lipid droplet dynamics using BODIPY staining, and for metabolic flux analysis with Seahorse technology. Functional assays including MTT proliferation, Transwell migration, Annexin V apoptosis, and drug sensitivity testing can elucidate ACSL5??s role in tumor cell behavior. Lipidomics profiling via LC-MS further enables detailed investigation of lipid remodeling. The polyclonal nature supports high-content applications. For additional technical information, contact Ascent Research.
