The GPT2 Knockout SK-HEP-1 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population of SK-HEP-1 cells in which the GPT2 gene has been disrupted, creating a loss-of-function model for studying mitochondrial alanine aminotransferase function. This polyclonal knockout cell population provides a heterogeneous pool of edited cells, suitable for pooled functional assays and metabolic characterizations without the selection bias of clonal isolates. It offers a ready-to-use system for investigating GPT2-mediated metabolic pathways in a human hepatic adenocarcinoma background.
SK-HEP-1 is a human cell line derived from the ascites of a 52-year-old male with hepatocellular carcinoma. This cell line exhibits a mixed phenotype with both epithelial and endothelial characteristics, and it is frequently employed as a model for liver sinusoidal endothelial cells. Its widespread use in liver cancer research and angiogenesis studies makes it a valuable host for investigating metabolic alterations in a tumor-relevant context. The SK-HEP-1 background supports the evaluation of gene function in cellular processes such as proliferation, migration, and metabolic adaptation.
GPT2 encodes mitochondrial alanine aminotransferase 2, which catalyzes the reversible conversion of alanine and 2-oxoglutarate to pyruvate and glutamate, dependent on the cofactor pyridoxal phosphate. Its expression is regulated by insulin, glucagon, glucocorticoids, and transcription factors HNF4A and FOXA2, as well as mTORC1 signaling. The pyruvate and glutamate produced feed into the TCA cycle and gluconeogenesis, directly linking amino acid metabolism to energy production and glucose synthesis. GPT2 cooperates with other aminotransferases such as GOT2 and GDH, and its activity influences mitochondrial function and cellular redox balance.
In the SK-HEP-1 hepatic adenocarcinoma model, disruption of GPT2 impairs the alanine?Cglucose metabolic axis, potentially attenuating gluconeogenesis and altering TCA cycle intermediate pools. Given the endothelial-like properties of this cell line, the knockout provides a unique platform to study how amino acid metabolism intersects with angiogenic signaling and tumor microenvironment interactions. This knockout model may reveal context-specific dependencies on GPT2 for proliferation, migration, and survival under nutrient-limited conditions.
Researchers can employ this polyclonal knockout product for metabolic flux analysis using LC?MS?based metabolomics or Seahorse extracellular flux assays, glucose production measurements, and amino acid tracing studies. It is suitable for investigating hepatocellular carcinoma metabolism, the role of GPT2 in gluconeogenesis, and the cellular response to hormonal regulators. Additional applications include cell migration and invasion assays, viability studies under metabolic stress, and mechanistic studies of neurodevelopmental disorders when coupled with relevant differentiation protocols. For additional technical details or to request a quotation, please contact Ascent Research.
