The ACOT11 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the ACOT11 gene has been disrupted to create a loss-of-function model for investigating mitochondrial fatty acid metabolism. This product consists of a heterogeneous pool of edited HEK293T cells, providing a robust and physiologically relevant system for studying the role of ACOT11 in energy homeostasis without the confounding effects of clonal selection. The polyclonal nature ensures that a broad spectrum of genetic disruptions is represented, enabling researchers to assess pooled phenotypic effects in downstream metabolic assays. As a ready-to-use knockout model, these cells serve as a critical tool for dissecting the molecular mechanisms linking fatty acyl-CoA hydrolysis to mitochondrial function.
HEK293T is a human embryonic kidney epithelial cell line immortalized through stable expression of the SV40 large T antigen, which confers high transfectability and ease of genetic manipulation. These adherent cells are widely employed as a versatile platform for gene-editing studies, protein expression, and signal transduction research. Their epithelial origin and robust growth characteristics make them particularly suited for transient and stable transfections, enabling efficient delivery of CRISPR components. In the context of metabolic research, HEK293T cells retain core fatty acid oxidation machinery, allowing meaningful interrogation of lipid metabolism pathways despite their non-hepatic lineage.
ACOT11 encodes a mitochondrial acyl-CoA thioesterase that catalyzes the hydrolysis of medium- and long-chain fatty acyl-CoAs to free fatty acids and coenzyme A within the mitochondrial matrix. This enzymatic activity is a key regulatory node in fatty acid catabolism, as it directly modulates the availability of substrates for ??-oxidation. ACOT11 is transcriptionally regulated by upstream factors, including PPAR?? and PGC-1??, and its expression is induced by fasting signals while being suppressed by insulin. Functionally, it acts downstream of these regulators to control free fatty acid pools and mitochondrial respiration. The enzyme interacts with acyl-CoA synthetases and CPT1, and its activity influences the expression of downstream effectors such as UCP1 and ??-oxidation enzymes, forming a critical link in the thermogenesis and insulin signaling pathways.
Disruption of ACOT11 in the HEK293T background creates a valuable model for exploring the consequences of impaired mitochondrial fatty acid handling. Although HEK293T cells are not canonical metabolic cells, they express key components of the fatty acid oxidation pathway, including CPT1A, ACOX1, and ACSL1, making them receptive to ACOT11-dependent metabolic shifts. Knockout of ACOT11 in this context is expected to alter mitochondrial respiration efficiency, fatty acid flux, and insulin signaling readouts, such as phospho-Akt levels. The polyclonal nature mitigates concerns about clonal artifacts and provides a more representative landscape of gene disruption, facilitating the identification of robust phenotypic changes in lipid accumulation and energy expenditure.
This knockout cell product is ideal for a variety of advanced research applications, including metabolic disease modeling of obesity, insulin resistance, and non-alcoholic fatty liver disease (NAFLD). Researchers can employ these cells in mechanistic studies of mitochondrial dysfunction, employing assays such as fatty acid oxidation measurements using etomoxir sensitivity, Seahorse-based mitochondrial respiration profiling, and triglyceride accumulation assays. Additionally, the cells are suitable for RNA-seq analysis to map global transcriptomic changes in metabolic gene expression, as well as drug screening efforts targeting metabolic syndrome. For further details on validation data and bulk pricing, please contact Ascent Research.
