The INSR Knockout 143B Polyclonal Cells constitute a human osteosarcoma-derived polyclonal knockout cell population generated via CRISPR/Cas9-mediated disruption of the INSR gene. This product provides a genetically mixed pool of edited cells, each carrying targeted disruptions at the insulin receptor locus, and serves as a robust loss-of-function model for investigating insulin and insulin-like growth factor (IGF) receptor signaling.
The parental 143B cell line is a well-characterized human osteosarcoma model originally derived from a bone tumor biopsy. 143B cells exhibit adherent, fibroblast-like morphology and are extensively used as a model system for bone cancer research. Their tumorigenic properties, rapid proliferation, and well-documented signaling pathways make them an ideal host for studying cancer cell metabolism, invasion, and response to therapeutic agents.
The INSR gene encodes the insulin receptor, a receptor tyrosine kinase that plays a central role in metabolic regulation and cellular growth. Upon binding of insulin, IGF1, or IGF2, the receptor undergoes autophosphorylation and recruits adaptor proteins including IRS1, IRS2, SHC, and GRB2. These interactions activate two major signaling cascades: the PI3K-AKT pathway and the MAPK/ERK pathway. Activated AKT promotes glucose uptake via GLUT4 translocation and modulates mTOR signaling, while ERK signaling influences cell proliferation and differentiation. Additionally, the receptor activity is modulated by the phosphatase PTP1B. In the absence of INSR, downstream phosphorylation of AKT and ERK is abrogated, disrupting insulin-dependent metabolic and mitogenic responses.
In the osteosarcoma background, INSR knockout cells provide a unique platform to dissect the contributions of insulin signaling to cancer cell biology. Osteosarcoma cells often rely on altered metabolic pathways to sustain rapid growth, and the INSR loss-of-function model allows researchers to uncouple insulin-driven anabolic signals from other oncogenic drivers. This is particularly relevant for studying the interplay between systemic insulin/IGF signals and bone tumor progression, as well as for modeling insulin resistance in a cancer context. Furthermore, these cells can be used to investigate compensatory mechanisms or signaling rewiring that may occur upon chronic INSR disruption.
Researchers can employ these polyclonal knockout cells in a wide array of functional assays. Insulin stimulation experiments followed by western blotting for phospho-AKT (Ser473) and phospho-ERK (Thr202/Tyr204) provide direct readouts of pathway inactivation. Glucose uptake assays using fluorescent or radiolabeled glucose analogues quantify metabolic defects. Cell proliferation and viability studies in the presence or absence of insulin enable assessment of growth factor dependency. RT-qPCR profiling of metabolic genes (e.g., GLUT4, HK2) can reveal transcriptional adaptations. Moreover, metabolic flux analysis and drug screening campaigns targeting the insulin/IGF axis can be conducted using this knockout model. For custom applications or further product details, please contact Ascent Research.
