The AKT1 Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited heterogeneous population of human 143B osteosarcoma cells harboring targeted disruptions in the AKT1 gene. This polyclonal format provides a diverse pool of loss-of-function alleles, minimizing clonal selection artifacts and enabling robust interrogation of AKT1-dependent signaling in a native cancer-cell context without the bias of monoclonal isolation.
The 143B cell line, a highly tumorigenic and metastatic subline of HOS derived from a 13-year-old female osteosarcoma patient, serves as a standard model for studying bone cancer progression, metastasis, and therapeutic responses. Its aggressive phenotype and reproducible in vivo tumor formation make it an ideal host for dissecting oncogenic kinase contributions.
AKT1 encodes a critical serine/threonine kinase within the PI3K/AKT pathway. Upon growth factor stimulation (EGF, IGF-1, PDGF) and receptor tyrosine kinase activation (EGFR, IGF1R, PDGFR), PI3K generates PIP3, recruiting AKT1 to the membrane where PDK1 and mTORC2 phosphorylate Thr308 and Ser473, respectively. Active AKT1 phosphorylates downstream effectors including GSK3??, FOXO1/3/4, mTORC1, BAD, and MDM2, regulating survival, proliferation, and metabolism. Negative regulation by PTEN, PP2A, and PHLPP, along with interactions with HSP90 and APPL1, finely tune AKT1 activity.
In 143B cells, AKT1 hyperactivation drives tumorigenic hallmarks such as apoptosis resistance, enhanced proliferation, and metastatic spread. Disruption of AKT1 attenuates phosphorylation of key substrates, reactivating pro-apoptotic and antiproliferative programs and compromising metabolic adaptation, thereby providing a powerful model to dissect AKT1-specific oncogenic mechanisms in osteosarcoma.
These polyclonal knockout cells are suited for phospho-kinase profiling, Western blotting, cell proliferation and apoptosis assays, migration and invasion studies, and drug sensitivity testing to validate AKT pathway inhibitors. Metabolic and rescue experiments further delineate isoform-specific functions. For additional information, please contact Ascent Research.
