The ACOT13 Knockout HEK293T Cell Line is a CRISPR/Cas9-mediated gene-disrupted cell line lacking functional ACOT13 expression. This product provides a defined loss-of-function model to study the role of acyl-CoA thioesterase 13 in fatty acid metabolism and intracellular acyl-CoA homeostasis. Engineered in the HEK293T background, it enables precise dissection of ACOT13-dependent lipid regulatory mechanisms without interference from endogenous protein.
HEK293T cells, derived from human embryonic kidney epithelium, constitutively express the SV40 large T antigen, which supports episomal replication of plasmids containing the SV40 origin of replication. This property confers high transfection efficiency and robust recombinant protein and viral vector production. The cell line??s well-characterized metabolic activity and genetic tractability make it a standard host for interrogating pathways governing lipid metabolism, protein expression, and signal transduction.
ACOT13 hydrolyzes medium- and long-chain fatty acyl-CoA thioesters into free fatty acids and CoA, thereby dampening intracellular acyl-CoA levels. Its expression is regulated by transcription factors PPARA, PPARG, SREBF1, and NR1H3, and is influenced by AMPK signaling. Loss of ACOT13 leads to accumulation of fatty acyl-CoAs, which can redirect metabolic flux away from mitochondrial ??-oxidation??mediated by CPT1A??and toward lipid synthesis via FASN, promoting triglyceride storage. This disruption also potentially affects ACSL1-mediated acyl-CoA formation and ACOX1-dependent peroxisomal oxidation, highlighting the enzyme??s integrative role in lipid homeostasis.
In the HEK293T context, ACOT13 knockout creates a versatile tool to model aspects of metabolic dysregulation observed in obesity, type 2 diabetes, NAFLD, and metabolic syndrome. Despite their non-hepatic origin, HEK293T cells retain core lipid handling machinery, allowing researchers to examine how altered acyl-CoA homeostasis influences lipid droplet dynamics, energy substrate utilization, and insulin signaling pathways. The knockout line is thus well-suited for mechanistic studies that require a genetically clean, easily manipulated background.
This cell line supports diverse experimental approaches: seahorse-based respirometry to quantify fatty acid oxidation, Oil Red O staining to visualize lipid droplets, LC-MS for precise acyl-CoA profiling, and immunoblotting or RT-qPCR to monitor metabolic enzyme adaptation. Applications include investigating acyl-CoA disequilibrium, screening small molecules that modulate lipid metabolism, and validating therapeutic targets for metabolic disorders. For additional information or to discuss custom services, please contact Ascent Research.
