The SOAT1 Knockout Hep-G2 Cell Line is a genetically modified derivative of the Hep-G2 hepatocellular carcinoma cell line, generated via CRISPR/Cas9-mediated disruption of the human SOAT1 gene. This knockout cell line provides a stable, renewable resource for investigating cholesterol esterification and lipid metabolism. By eliminating SOAT1 activity, researchers can directly dissect its role in hepatocellular lipid homeostasis without off-target effects associated with chemical inhibitors.
Hep-G2 originates from a 15-year-old male hepatocellular carcinoma patient and is a well-characterized hepatic cell model. It retains many hepatocyte-specific functions, including drug-metabolizing enzyme activities, albumin secretion, and regulated lipid metabolism, making it a preferred platform for hepatotoxicity testing and metabolic research. In this SOAT1 knockout derivative, the hepatic background is preserved, allowing study of gene function in a pathophysiologically relevant context.
SOAT1 encodes sterol O-acyltransferase, an endoplasmic reticulum-resident enzyme that catalyzes the conversion of free cholesterol to cholesteryl esters for storage in lipid droplets. Its expression is transcriptionally activated by SREBP2 and LXR in response to cellular sterol status, and its enzymatic activity is further modulated by insulin signaling pathways. SOAT1 physically interacts with the lipid droplet coat proteins PLIN2 and PLIN3 and cooperates with ACAT2 to mediate cholesteryl ester deposition. Downstream, SOAT1 activity reduces membrane free cholesterol levels, thereby promoting ABCA1- and ABCG1-dependent cholesterol efflux and influencing ApoB-mediated VLDL secretion. This positions SOAT1 at a central juncture of cholesterol metabolism, lipid droplet biogenesis, and lipoprotein assembly, functionally interconnected with HMGCR and LDLR.
Disruption of SOAT1 in Hep-G2 cells perturbs hepatic cholesterol homeostasis, leading to diminished cholesteryl ester accumulation, aberrant lipid droplet morphology, and altered VLDL secretion profiles. These phenotypic changes recapitulate key features of human lipid storage disorders and provide a cell-based model for non-alcoholic fatty liver disease (NAFLD), hypercholesterolemia, and the metabolic underpinnings of hepatocellular carcinoma. The knockout line enables detailed mechanistic studies of SOAT1’s role in hepatocyte lipid partitioning and its contribution to disease progression, bridging molecular insights with translational applications.
This SOAT1 knockout model is well-suited for a range of functional assays, including [14C]oleate incorporation to measure cholesterol esterification, Oil Red O and BODIPY staining for lipid droplet visualization, LDL uptake and cholesterol efflux assays, and qRT-PCR or western blotting for pathway analysis. It supports target validation and compound screening in atherosclerosis, NAFLD, and hyperlipidemia drug discovery. The cell line serves as a renewable genetic tool for basic and preclinical research. For technical inquiries, please contact Ascent Research.
