The GSK3A Knockout 143B Polyclonal Cells product is a pool of CRISPR/Cas9-edited polyclonal knockout cells generated from the 143B human osteosarcoma line, with disruption of the GSK3A gene. This polyclonal population provides a versatile loss-of-function model for studying GSK3A function in a bone cancer context, avoiding clonal artifacts while maintaining effective gene knockout.
The 143B cell line is a highly tumorigenic human osteosarcoma model widely employed in cancer research. It is particularly valuable for investigating bone tumor biology, signaling mechanisms, and therapeutic responses due to its aggressive growth and relevance to osteosarcoma pathogenesis.
GSK3A encodes an isoform of glycogen synthase kinase-3 that phosphorylates key substrates such as glycogen synthase (GYS1), ??-catenin (CTNNB1), and tau (MAPT). In Wnt/??-catenin signaling, GSK3A operates in the destruction complex with Axin, APC, and ??-catenin, promoting ??-catenin degradation; Wnt ligands via Frizzled/LRP5/6 and Dishevelled inhibit GSK3A, stabilizing ??-catenin for TCF/LEF-mediated transcription of c-Myc and cyclin D1. Insulin/IGF-1 receptors activate PI3K/AKT, which phosphorylate GSK3A at Ser21, reducing its activity and impacting glycogen metabolism and mTOR signaling. Interacting partners include FRAT1 and AKT, and pathways such as insulin, PI3K/AKT, and mTOR converge on GSK3A to regulate cell proliferation, survival, and metabolism.
In 143B osteosarcoma cells, GSK3A knockout eliminates inhibitory phosphorylation of downstream targets, leading to constitutive activation of Wnt/??-catenin and AKT signaling cascades. This amplifies ??-catenin-dependent transcription and prosurvival signals, driving proliferation, metabolic alterations, and apoptosis resistance. The model therefore enables dissection of GSK3A-specific roles in osteosarcoma without GSK3B interference.
Applications include Wnt pathway analysis via ??-catenin/TCF reporter assays, proliferation and apoptosis studies with MTS/MTT and Annexin V, and cell cycle profiling by flow cytometry. Knockout confirmation and downstream effects can be assessed by western blotting for GSK3A, phospho-AKT, and phospho-substrates, or by RT-qPCR. Transcriptomic analysis by RNA-seq further supports target discovery. This product is suited for drug target validation of GSK3 inhibitors in cancer metabolism. For further details, please contact Ascent Research.
