The IGF2 Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the 143B human osteosarcoma cell line, featuring targeted disruption of the IGF2 gene. This polyclonal knockout allows study of insulin-like growth factor 2 function in a bone cancer context without single-cell cloning bias.
The 143B cell line is a widely employed human osteosarcoma model derived from a primary bone tumor biopsy. Characterized by rapid growth, high invasiveness, and metastatic potential, this line serves as a standard platform for bone cancer research, including investigations into tumorigenesis, drug response, and signaling networks.
IGF2 is a potent growth factor that binds IGF1R to activate PI3K-Akt signaling via IRS1, leading to AKT-mediated phosphorylation of mTOR, S6K, and FOXO1, thereby promoting protein synthesis, cell cycle progression, and survival. Concurrently, IGF1R engages the SHC-GRB2-SOS complex, triggering RAS-RAF-MEK-ERK cascade and upregulating targets such as cyclin D1 and c-Myc, which drive proliferation. IGF2 bioavailability is influenced by interactions with IGFBP3, IGFBP5, and the clearance receptor IGF2R, and its expression is transcriptionally regulated by PLAG1, PLAGL1, and p53, with additional control by epigenetic imprinting.
In 143B osteosarcoma cells, IGF2 commonly functions as an autocrine factor sustaining constitutive activation of AKT and ERK pathways, contributing to uncontrolled proliferation and resistance to apoptosis. CRISPR/Cas9-mediated disruption of IGF2 abrogates this signaling, leading to reduced phosphorylation of AKT and ERK, decreased expression of downstream effectors such as cyclin D1 and Bcl-2, and impaired cell cycle progression and colony formation. This knockout model thus enables rigorous dissection of IGF2-specific contributions to osteosarcoma phenotype and evaluation of signaling dependency.
Typical applications include cell proliferation assays (MTT or BrdU incorporation), apoptosis detection by Annexin V staining, and cell cycle profiling by flow cytometry to quantify growth and death responses. Clonogenic survival can be assessed through colony formation, while migration and invasion assays interrogate metastatic behavior. Molecular characterization involves western blotting and RT-qPCR to confirm IGF2 protein and mRNA levels, along with phospho-specific antibodies for AKT and ERK to monitor pathway activity. The polyclonal population is also amenable to RNA-seq for genome-wide expression analysis. For comprehensive technical details and ordering, please contact Ascent Research.
