GOT1 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population originated from the human liver hepatocellular carcinoma cell line SK-HEP-1. The product features a targeted disruption of the GOT1 gene using CRISPR/Cas9 technology, generating a heterogeneous pool of cells with loss of GOT1 function. This polyclonal format ensures that experimental outcomes reflect average population effects, avoiding the bias of single-cell clones and providing a dependable model for interrogating GOT1-related biology.
The SK-HEP-1 parental line was established from the ascites of a patient with liver adenocarcinoma and is a classic model for hepatocellular carcinoma research. These cells display an epithelial phenotype with mesenchymal features and are extensively employed to study hepatic tumor biology, metastatic dissemination, and drug metabolism. Their well-characterized metabolic profile makes them particularly appropriate for examining the impact of GOT1 disruption on cancer cell metabolism.
GOT1 encodes cytosolic aspartate aminotransferase, an enzyme that catalyzes the reversible conversion of aspartate and alpha-ketoglutarate to glutamate and oxaloacetate, dependent on the pyridoxal phosphate cofactor. As a core component of the malate-aspartate shuttle, GOT1 cooperates with MDH1 and the mitochondrial carrier SLC25A13 to transfer reducing equivalents across the inner mitochondrial membrane, thereby sustaining the cytosolic NAD+/NADH ratio. The gene is transcriptionally activated by oncogenic factors such as c-MYC, HIF-1??, and NRF2. Disruption of GOT1 disrupts this shuttle, blunts oxaloacetate regeneration, cripples aspartate synthesis, curtails nucleotide biosynthesis, and erodes redox homeostasis, ultimately impairing cellular proliferation and resilience to oxidative stress.
Within the SK-HEP-1 context, GOT1 knockout imposes a severe metabolic constraint that limits aspartate-dependent anabolic processes and disturbs redox balance. This renders the cells more reliant on alternative nutrient sources and sensitizes them to insults that target antioxidant defenses. The model therefore offers a valuable system for dissecting how hepatocellular carcinoma cells adapt to aspartate scarcity and for evaluating the therapeutic potential of strategies that exploit redox vulnerabilities in liver tumors.
The GOT1 Knockout SK-HEP-1 Polyclonal Cells support a broad range of experimental workflows, including metabolomic profiling via LC-MS, real-time PCR and western blotting for expression analysis, NAD+/NADH quantification, oxygen consumption rate (OCR) measurement, and functional assays such as proliferation, apoptosis, and clonogenicity tests. They are particularly suited for synthetic lethality screens targeting redox regulation and for validating metabolic drug targets in hepatocellular carcinoma. For further details, please contact Ascent Research.
