The BRAF Knockout 143B Polyclonal Cells product comprises a heterogeneous population of 143B human osteosarcoma cells subjected to CRISPR/Cas9-mediated disruption of the BRAF gene. This polyclonal knockout format provides a pool of cells harboring diverse loss-of-function mutations within the BRAF locus, avoiding clonal selection biases and enabling bulk functional studies in a cancer-relevant background. The resulting cell model serves as a valuable tool for dissecting BRAF-dependent signaling networks and assessing pathway dependencies in a bone tumor setting.
The parental 143B cell line originates from a human osteosarcoma, a primary malignant bone tumor characterized by osteoblastic features. This adherent cell line is widely employed as a model for investigating osteosarcoma pathogenesis, bone metastasis, and tumor?Cbone microenvironment interactions. Its robust growth properties and genetic background make it suitable for downstream assays addressing skeletal malignancies and therapeutic responses.
BRAF is a serine/threonine kinase that transmits signals from activated RAS (KRAS, HRAS, NRAS) to the MEK?CERK cascade. Activated BRAF phosphorylates MEK1 (MAP2K1) and MEK2 (MAP2K2), which then phosphorylate ERK1 (MAPK3) and ERK2 (MAPK1). Nuclear ERK1/2 regulates transcription factors including ELK1, MYC, FOS, and JUN, thereby modulating proliferation, survival, and differentiation. Its activity is modulated by upstream receptors (EGFR, FGFR), kinases (SRC, PKA), and scaffold proteins (KSR), and it interacts with 14?3?3 and HSP90 for proper folding and signal fidelity. This BRAF?CMEK?CERK axis is a central node in the RAS?CMAPK pathway, frequently dysregulated in cancer.
In the 143B osteosarcoma context, although BRAF mutations are not typical, the MAPK pathway is frequently hyperactivated through other mechanisms, such as growth factor receptor overexpression or RAS mutations. Disruption of BRAF in these cells allows researchers to probe the necessity of intact MAPK flux for osteosarcoma cell proliferation, survival, migration, and drug resistance. Moreover, this model is instrumental for studying crosstalk between the MAPK pathway and bone?remodeling signals, contributing to understanding of tumor?Cbone interactions in metastatic or primary bone malignancies.
This polyclonal knockout population supports a wide range of assays, including western blotting for phospho?MEK and phospho?ERK to confirm pathway abrogation, RT?qPCR for downstream targets such as CCND1 and MYC, cell proliferation (MTT, BrdU), migration/invasion assays, and drug sensitivity testing using BRAF inhibitors (vemurafenib, dabrafenib). Flow cytometry enables assessment of cell cycle distribution and apoptosis. The model is suited for phenotypic screens aiming to identify synthetic lethal interactions or resistance mechanisms to MAPK?directed therapies. For further technical specifications and ordering information, please contact Ascent Research.
