The ACOT7 Knockout HT29 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, featuring disruption of the ACOT7 gene. This loss-of-function model enables the study of ACOT7’s role in lipid metabolism and associated signaling pathways. The polyclonal population contains a heterogeneous mix of cells with targeted gene disruption, making it suitable for pooled functional studies, drug screening, and biochemical analyses without clonal selection artifacts.
HT29 cells are a well-established adherent epithelial cell line originating from a primary colorectal adenocarcinoma of a 44-year-old female. These cells retain some differentiated intestinal epithelial characteristics, including the capacity for enterocytic differentiation, and maintain tumorigenic properties. As a model of colorectal cancer, HT29 cells are widely used for investigating cancer cell biology, metabolism, and therapeutic responses, providing a relevant context for examining the consequences of ACOT7 knockout in a transformed intestinal epithelial background.
ACOT7 (acyl-CoA thioesterase 7) hydrolyzes medium- and long-chain fatty acyl-CoAs to free fatty acids and coenzyme A, thereby modulating intracellular acyl-CoA pools and influencing fatty acid oxidation and lipid signaling. The enzyme is regulated by PPAR?? and PPAR?? transcription factors, insulin, and fatty acids, and its activity is interconnected with acyl-CoA synthetases (ACSLs) and peroxisomal import via PEX5. ACOT7-mediated production of free fatty acids can affect downstream PPAR target genes such as CPT1A and ACOX1, as well as the synthesis of inflammatory mediators like prostaglandins, positioning ACOT7 at a critical node linking lipid metabolism, mitochondrial function, and inflammatory signaling.
In the HT29 colorectal cancer context, ACOT7 disruption is expected to alter lipid homeostasis and energy metabolism, potentially impacting the balance between fatty acid ??-oxidation and lipid storage. Given the dependence of many cancer cells on lipid metabolism for membrane biosynthesis and energy, ACOT7 knockout may influence cell proliferation, differentiation, and sensitivity to metabolic stress. This model is therefore valuable for exploring how acyl-CoA thioesterase activity contributes to colorectal cancer phenotypes and for investigating mechanisms relevant to metabolic disorders and spastic paraplegia 41, a disease associated with ACOT7 mutations.
Applications include but are not limited to lipidomics, fatty acid oxidation assays, and Seahorse metabolic flux analysis to assess mitochondrial and glycolytic function. The polyclonal ACOT7 knockout cells can be employed in PPAR reporter assays, RT-qPCR and western blotting of metabolic enzymes, immunofluorescence imaging of peroxisomal markers, and cell proliferation or drug sensitivity testing. This product is suitable for cancer metabolism research, peroxisomal biology studies, and drug screening aimed at metabolic targets. For detailed protocol recommendations or to inquire about pricing and availability, please contact Ascent Research.
