The ACOT8 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the ACOT8 gene in the HeLa human cervical carcinoma cell line. This loss-of-function model supports detailed investigations into acyl-CoA thioesterase 8, an enzyme critical for intracellular fatty acyl-CoA hydrolysis. The polyclonal format offers a heterogeneous knockout pool, minimizing clonal selection artifacts and enabling robust, reproducible phenotypic analyses. Researchers can systematically evaluate the consequences of ACOT8 disruption on lipid metabolism in a widely used epithelial cancer model.
HeLa cells, derived from a cervical adenocarcinoma, are immortalized by HPV18-mediated inactivation of p53 and retinoblastoma protein. This oncogenic transformation grants rapid proliferation, aneuploidy, and a pronounced capacity for lipid metabolic reprogramming. As a well-characterized model in cancer biology, HeLa provides an excellent platform for dissecting gene functions linked to peroxisomal biology, lipid signaling, and metabolic rewiring. The host??s extensive molecular toolkit and existing datasets further enhance its utility for CRISPR-based knockout studies.
ACOT8 encodes a peroxisomal and cytosolic thioesterase that hydrolyzes medium- to long-chain fatty acyl-CoAs into free fatty acids and CoA. ACOT8 expression is upregulated by PPAR?? and SREBP1c and is modulated by insulin and fatty acid levels. The released CoA contributes to acetyl-CoA and malonyl-CoA pools, influencing mitochondrial and peroxisomal beta-oxidation as well as lipid synthesis. ACOT8 interacts with peroxisomal import receptors PEX5 and PEX7 and acyl-CoA binding proteins. In the metabolic network, ACOT8 cooperates with enzymes such as ACOX1 and CPT1A to govern fatty acid flux and energy balance.
Knockout of ACOT8 in HeLa cells disrupts the acyl-CoA/free fatty acid equilibrium, providing a system to probe lipid metabolic vulnerabilities in cancer. The HeLa background??s high proliferation rate places unique demands on lipid supply and CoA recycling; ACOT8 loss may uncover dependencies in peroxisomal beta-oxidation and membrane biogenesis. This model is particularly suited to explore how altered acyl-CoA thioesterase activity affects lipid droplet dynamics and oncogenic signaling. It enables dissection of ACOT8-dependent pathways in an HPV-transformed, aneuploid context.
Applications include LC-MS-based fatty acid profiling, CoA quantification, beta-oxidation flux assays, and lipid droplet staining. Proliferation assays (MTS/MTT) and metabolic tracing with labeled fatty acids reveal the impact on cell growth and energy metabolism. The polyclonal format is ideal for pooled genetic screens and dose-response drug studies targeting lipid metabolism. For further technical details or to place an order, please contact Ascent Research.
