The ACOT7 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa human cervical adenocarcinoma cell line. This product features targeted disruption of the ACOT7 gene, which encodes acyl-CoA thioesterase 7, an enzyme that hydrolyzes medium- to long-chain fatty acyl-CoAs to free fatty acids and coenzyme A. The polyclonal nature of this knockout pool ensures representation of diverse editing events across the cell population, providing a robust loss-of-function model for studying ACOT7-dependent lipid metabolism without requiring clonal selection. The cells are supplied as a heterogeneous mixture, reflecting a range of editing outcomes, and are suitable for experiments where population-level analysis is preferred over monoclonal characterization.
The host HeLa cell line is an extensively characterized model of human cervical epithelial adenocarcinoma, originally derived from a patient with HPV18-positive cervical carcinoma. HeLa cells exhibit a transformed phenotype with high proliferation rate, aneuploidy, and documented metabolic alterations, making them a widely used system for investigating cancer cell biology and metabolic reprogramming. Their human origin and epithelial characteristics provide a physiologically relevant context for studying ACOT7 function in a cancer setting, while the HPV18 status adds a layer of oncogenic background relevant to viral-driven metabolic changes.
ACOT7 functions as a key regulator of intracellular acyl-CoA levels and fatty acid metabolism by catalyzing the hydrolysis of fatty acyl-CoAs, thereby releasing free fatty acids and CoA. This activity influences the availability of acyl-CoAs for ??-oxidation, membrane lipid synthesis, and signaling molecule production. The enzyme is regulated by PPAR??, insulin, and fatty acids, and interacts with HNF4?? and fatty acid-binding protein 1 (FABP1). Downstream, ACOT7 activity modulates the CoA pool and promotes PPAR??-mediated transcriptional programs, impacting genes such as ACOX1, CPT1A, and SCD1. Disruption of ACOT7 is expected to cause accumulation of long-chain acyl-CoAs, which can allosterically affect metabolic enzymes and transcription factors, thereby altering lipid metabolism and energy homeostasis.
In the HeLa cellular context, ACOT7 knockout presents a unique model to examine the intersection of lipid metabolism and cancer cell physiology. HeLa cells rely on balanced fatty acid flux for membrane biogenesis, energy production, and signaling. Loss of ACOT7-mediated hydrolysis disrupts this balance, potentially leading to acyl-CoA accumulation that may feedback-inhibit fatty acid oxidation or alter lipid droplet dynamics. These changes can shift cellular reliance toward alternative metabolic pathways, influence PPAR?? target gene expression, and modify membrane lipid composition, which in turn may affect proliferation, survival, and tumorigenic properties. The HPV18-positive background further suggests potential interplay between viral oncoproteins and lipid metabolic reprogramming, making this model valuable for studying metabolic vulnerabilities in cervical carcinoma.
These polyclonal knockout cells are suitable for a broad range of research applications, including lipid metabolism studies, PPAR signaling investigation, cancer metabolism research, and metabolic disease modeling. Researchers can employ assays such as acyl-CoA thioesterase activity measurements, LC-MS?Cbased lipidomics, RT-qPCR for ACOT7 and PPAR target genes, cellular free fatty acid quantification, Western blotting for PPAR??, Seahorse metabolic flux analysis for oxygen consumption and extracellular acidification rates, and MTT cell viability assays. This model facilitates drug target validation for metabolic disorders like obesity and type 2 diabetes, as well as exploration of neurodegenerative disease mechanisms. For technical specifications, batch validation data, or ordering inquiries, please contact Ascent Research.
