The GYS1 Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the glycogen synthase 1 (GYS1) gene in the human 143B osteosarcoma cell line. Supplied as a heterogeneous pool of cells with disrupted GYS1 expression, this product serves as a loss-of-function model for investigating glycogen metabolism and metabolic signaling in an osteoblast-like bone cancer context.
Established from a human osteosarcoma, the 143B cell line exhibits an osteoblast-like phenotype and is widely used in bone metastasis and cancer metabolism studies. These adherent cells retain signaling pathways relevant to osteosarcoma biology and energy homeostasis, providing a clinically relevant background for examining the interplay between glycogen storage and tumor cell adaptation.
GYS1 encodes the rate-limiting enzyme of glycogen synthesis, catalyzing glucose transfer from UDP-glucose to glycogen. Its activity is regulated by insulin-induced AKT activation, which inhibits GSK3??, allowing PP1 complexes containing PPP1R3 to dephosphorylate and activate GYS1. Conversely, energy stress signals through AMPK and PKA phosphorylate and inactivate GYS1. Downstream, GYS1 controls glycogen production and glucose-6-phosphate levels for glycolysis. Key interacting factors include glycogenin (GYG1) and PPP1CA.
Knockout of GYS1 in 143B cells abrogates glycogen synthesis, profoundly reshaping energy metabolism and glucose utilization. This elimination forces cells to rely on alternative carbon sources, altering their response to metabolic stress and mimicking aspects of glycogen storage disease type 0. In the osteosarcoma context, GYS1 disruption uncovers vulnerabilities in tumor metabolism, particularly under nutrient deprivation or during metastasis, where glycogen reserves often support survival.
This polyclonal knockout pool is suited for glycogen content analyses via PAS staining, Western blotting for GYS1 and phospho-GYS1, glucose uptake measurements with 2-NBDG, and metabolic flux analyses using Seahorse analyzers. Applications include investigations of energy stress responses, insulin signaling disruptions, and osteosarcoma metabolic adaptations. The polyclonal format is particularly useful for viability assays under nutrient deprivation, capturing population-level variability. For additional information or technical inquiries, please contact Ascent Research.
