The GYS1 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the SK-HEP-1 human hepatic adenocarcinoma cell line. This product comprises a heterogeneous pool of cells harboring targeted disruption of the GYS1 gene, leading to loss of glycogen synthase 1 function. The polyclonal format avoids clonal selection bias, retaining metabolic heterogeneity for bulk-population studies of glycogen synthase deficiency. CRISPR/Cas9-mediated gene disruption generates a loss-of-function model, enabling investigation of GYS1 ablation in a liver cancer context without clonal isolation.
The SK-HEP-1 host cell line is an epithelial line established from ascites fluid of a hepatocellular carcinoma patient. It is extensively used as a model for liver cancer biology, including studies of metastasis, drug metabolism, and signaling networks. These cells are well-suited for examining metabolic adaptations in hepatic tumors and for drug screening applications, owing to their hepatic origin and robust in vitro growth characteristics.
GYS1 encodes glycogen synthase 1, the rate-limiting enzyme of glycogenesis, catalyzing glucose transfer from UDP-glucose to glycogen. It acts downstream of insulin signaling, where Akt-mediated phosphorylation inhibits GSK-3??, relieving inhibitory phosphorylation on GYS1. Protein phosphatase 1 (PP1), complexed with regulatory subunits (PPP1R3), dephosphorylates and activates GYS1, while AMPK phosphorylates and inhibits it under energy stress. GYS1 interacts with glycogenin to initiate glycogen particle assembly and is allosterically activated by glucose-6-phosphate. This signaling module controls cellular glycogen stores and glycolytic intermediate levels, connecting insulin action to glucose homeostasis. Dysregulation contributes to glycogen storage disease type 0, type 2 diabetes, and hepatocellular carcinoma.
Ablation of GYS1 in SK-HEP-1 cells abolishes glycogen synthase activity, preventing glycogen accumulation. This disrupts energy storage and alters metabolic signaling, potentially affecting proliferation and stress responses in hepatic cancer cells. The knockout model is valuable for dissecting how glycogen synthesis supports tumor survival under nutrient stress or hypoxia and for evaluating drug sensitivities linked to glycogen dependency. Loss of GYS1 provides insight into metabolic vulnerabilities in liver cancer.
Researchers can employ these cells to study glycogen metabolism in liver cancer using PAS staining, glycogen quantification, and metabolic flux analysis. The polyclonal knockout population is suitable for testing anti-cancer therapies that target glycogenesis, analyzing insulin signaling via phospho-Akt/GSK-3?? immunoblotting, and assessing proliferation under varied glucose conditions. Western blotting and RT-qPCR confirm GYS1 disruption. These cells are ideal for metabolic reprogramming studies and screening compounds that exploit glycogen dependency. For more information or custom quotes, contact Ascent Research.
