The AACS Knockout SK-HEP-1 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human SK-HEP-1 cell line, designed for targeted disruption of the AACS gene. This population consists of a heterogeneous pool of edited cells, faithfully representing the genetic and phenotypic variability inherent to polyclonal knockout models without the bias of clonal selection. By abolishing AACS function across the population, the model enables physiologically relevant studies of ketone body metabolism and its integration with lipid synthesis pathways.
SK-HEP-1 is a well-characterized human hepatic adenocarcinoma cell line exhibiting epithelial morphology, widely employed as a model system for hepatocellular carcinoma (HCC) and liver metabolism research. These cells recapitulate key metabolic and oncogenic features of liver cancer, including dysregulated lipid and energy homeostasis, making them an appropriate host for investigating the role of ketone body utilization in tumor biology. The SK-HEP-1 background provides a robust and reproducible platform for dissecting metabolic reprogramming mechanisms in HCC.
AACS encodes acetoacetyl-CoA synthetase, which catalyzes the cytoplasmic activation of acetoacetate to acetoacetyl-CoA, a critical step in ketone body catabolism. This reaction directly links ketone body utilization to the mevalonate pathway and fatty acid synthesis, as acetoacetyl-CoA serves as a substrate for HMG-CoA synthase and acetyl-CoA carboxylase. AACS transcription is regulated by SREBP transcription factors and insulin/glucagon signaling, positioning the enzyme as a nutrient-sensitive node connecting ketone metabolism to cholesterol biosynthesis and fatty acid elongation. Additionally, AACS interacts with acetoacetyl-CoA thiolase and other metabolic enzymes in the lipogenic cascade.
Knockout of AACS in SK-HEP-1 cells abrogates cytoplasmic acetoacetyl-CoA production, thereby impairing the synthesis of cholesterol and fatty acids from ketone body precursors. This metabolic disruption can alter membrane lipid composition, perturb energy homeostasis, and potentially sensitize HCC cells to metabolic stress, revealing vulnerabilities that depend on ketone body-fueled lipogenesis. Consequently, the model offers a powerful tool for examining how ketone body utilization supports tumor cell growth and survival in the context of liver cancer.
These polyclonal knockout cells are suited for a wide range of functional studies, including metabolic flux analysis using isotopically labeled acetoacetate, liquid chromatography?Cmass spectrometry (LC-MS) quantification of acetoacetyl-CoA, Oil Red O staining for neutral lipid assessment, cell proliferation assays under varied nutritional conditions, and gene expression analysis of cholesterol synthesis markers via RT-qPCR or western blot for HMG-CoA synthase. Applications extend to drug target validation and exploration of metabolic syndrome-related pathways. For further information, please contact Ascent Research.
