The ACOT11 Knockout HeLa Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal cell population derived from HeLa cells, in which the ACOT11 gene has been disrupted. This product provides a versatile loss-of-function model for studying the biological roles of ACOT11, a key enzyme in lipid metabolism, within an established epithelial cell context.
HeLa cells are an immortalized cervical adenocarcinoma line positive for HPV18, whose E6 and E7 oncoproteins inactivate p53 and Rb tumor suppressors, respectively, creating a hyperproliferative state. These adherent epithelial cells are extensively used in cancer research, virology, and cell biology, and their robust metabolic adaptability makes them ideal for metabolic studies.
ACOT11 encodes a long-chain fatty acyl-CoA thioesterase that hydrolyzes acyl-CoAs to free fatty acids and CoA, regulating intracellular fatty acid levels. It integrates into fatty acid metabolism, PPAR signaling, AMPK signaling, and insulin signaling pathways. Transcriptionally controlled by PPAR?? and SREBP1c, its enzymatic activity is modulated by insulin, glucagon, and AMPK. Downstream, it governs free fatty acid availability for mitochondrial oxidation and lipogenesis, influencing CoA and lipid second messengers. ACOT11 interacts with acyl-CoA synthetases and CPT1. Knockout results in acyl-CoA accumulation, disrupting the oxidation/synthesis balance and affecting PPAR?? and AMPK-dependent signaling networks.
In HeLa cells, the HPV-driven p53/Rb inactivation creates a hyperproliferative background; ACOT11 knockout further perturbs lipid homeostasis, potentially revealing metabolic vulnerabilities in cancer. This model is relevant to metabolic syndrome, type 2 diabetes, obesity, NAFLD, and cancer metabolism, where dysregulated fatty acid handling is pivotal. The polyclonal population reflects heterogeneous gene disruption, enabling study of functional consequences across a range of editing events.
These knockout cells are suited for fatty acid oxidation assays, lipidomics by mass spectrometry, and CoA quantification via LC-MS. Gene expression and protein analysis by RT-qPCR and western blotting can verify ACOT11 loss and downstream targets like CPT1A or ACOX1. Metabolic flux analysis using Seahorse respirometry and Oil Red O staining further define the lipid phenotype. Applications extend to drug target validation and compound screening. For further information, please contact Ascent Research.
