The JAG1 Knockout 143B Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population from the 143B osteosarcoma line, with targeted disruption of the JAG1 gene. This live cell product provides a genetically diverse loss-of-function model for investigating Jagged1-dependent Notch signaling in bone cancer, avoiding clonal artifacts and suited for direct culture and functional assays.
The 143B cell line is a TP53-deficient osteosarcoma line derived from HOS, widely employed to study osteosarcoma tumorigenesis, metastasis, and stromal interactions. Its TP53-null background mimics a frequent genetic lesion in high-grade osteosarcomas, offering a clinically relevant context for dissecting signaling pathways that drive tumor progression, including the Notch cascade.
Jagged1 is a transmembrane ligand that activates Notch receptors (NOTCH1, NOTCH2) on neighboring cells, initiating ADAM10/ADAM17-mediated cleavage and gamma-secretase-dependent release of the Notch intracellular domain (NICD). NICD translocates to the nucleus and forms a transcriptional complex with RBPJ and MAML coactivators to induce expression of effectors such as HES1, HEY1, MYC, and CCND1. JAG1 is regulated by TP53, HIF1??, and TGF-??1, and its signaling intersects with Wnt and PI3K/AKT pathways to control cell fate, proliferation, and survival. CRISPR/Cas9-mediated knockout eliminates JAG1, abrogating ligand-dependent Notch activation for unambiguous pathway analysis.
In the 143B osteosarcoma background, JAG1 knockout enables systematic study of Notch-driven processes such as proliferation, migration, invasion, and chemoresistance. This model is particularly suited for examining Jagged1-mediated paracrine communication with osteoblasts, endothelial cells, and immune components, which contributes to the metastatic niche. The TP53-deficient context also allows exploration of the cooperation between p53 loss and Notch signaling in aggressive osteosarcoma phenotypes.
Standard techniques include RT-qPCR for Notch target genes (HES1, HEY1), Notch luciferase reporter assays, and flow cytometry for NOTCH1 surface levels. Co-culture with Notch reporter cells measures Jagged1 trans-activation, while Transwell and MTS assays quantify migration and proliferation impacts. Apoptosis can be evaluated via Annexin V staining. The polyclonal knockout cells are well-suited for Notch inhibitor testing and high-content screens to uncover Jagged1-dependent vulnerabilities. For technical inquiries, contact Ascent Research.
