ACOT2 Knockout HT29 Polyclonal Cells offer a CRISPR/Cas9-edited polyclonal cell population derived from the human colorectal adenocarcinoma line HT29, featuring targeted disruption of the ACOT2 gene to create a robust loss-of-function model. This heterogeneous knockout pool enables functional studies without the confounding effects of clonal selection, providing a physiologically relevant system for investigating peroxisomal lipid metabolism.
The HT29 host cell line exhibits epithelial morphology and is widely utilized as a model for intestinal barrier function, drug transport, and colorectal cancer research. Its adenocarcinoma origin makes it particularly suited for exploring the intersection of metabolic reprogramming and tumor biology, with the ACOT2 knockout enabling direct examination of lipid metabolism in a cancer context.
ACOT2 encodes a peroxisomal acyl-CoA thioesterase that hydrolyzes long-chain acyl-CoAs into free fatty acids and CoA, a reaction essential for regulating peroxisomal fatty acid ??-oxidation and maintaining cellular CoA homeostasis. ACOT2 expression is activated downstream of PPAR?? and PPAR?? nuclear receptors, and the enzyme interacts with the peroxisomal import receptor PEX5. Its activity feeds into the broader ??-oxidation pathway, intersecting with ACOX1 and EHHADH, and modulates the availability of free fatty acids for lipid mediator synthesis and PPAR ligand generation.
In HT29 colorectal adenocarcinoma cells, ACOT2 knockout disrupts peroxisomal lipid handling, likely altering fatty acid oxidation rates and CoA balance. This metabolic perturbation can impinge upon PPAR signaling and free fatty acid?Cdependent pathways, offering a valuable tool to dissect how lipid metabolic flexibility supports colorectal cancer cell proliferation and survival.
Researchers can employ these polyclonal knockout cells in diverse assays including fatty acid ??-oxidation measurement, western blotting and RT-qPCR to confirm ACOT2 loss and downstream effects, immunofluorescence for peroxisomal markers such as PMP70, lipid profiling to assess free fatty acid shifts, and cell proliferation assays. The model is also suitable for screening inhibitors that target lipid metabolism in cancer. For more information or to discuss custom applications, please contact Ascent Research.
