The INSR Knockout 769-P Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population generated from the 769-P human clear cell renal cell carcinoma (ccRCC) line, featuring targeted disruption of the insulin receptor gene (INSR). This product comprises a heterogeneous pool of cells carrying diverse loss-of-function alleles, avoiding clonal selection bias and enabling robust population-level analyses of insulin signaling ablation. The polyclonal format is particularly suitable for dose?Cresponse studies, drug screening, and other experiments that benefit from averaged biological responses.
The 769-P cell line is a widely used model of ccRCC, derived from a primary tumor and harboring a biallelic loss-of-function mutation in the von Hippel?CLindau (VHL) tumor suppressor gene. VHL deficiency leads to constitutive stabilization of hypoxia-inducible factors (HIFs), driving a pseudohypoxic state that reprograms central carbon metabolism toward aerobic glycolysis and promotes angiogenesis. The cells exhibit an adherent epithelial morphology and are well-characterized for investigating VHL-dependent signaling and metabolic adaptation in kidney cancer.
The INSR gene product is a receptor tyrosine kinase activated by insulin, IGF-1, and IGF-2. Ligand binding triggers autophosphorylation and recruitment of IRS1/2 and SHC adaptors. IRS1/2 engage PI3K (catalytic p110 and regulatory p85/PIK3R1) to generate PIP3, activating AKT and downstream mTORC1. SHC?CGRB2?CSOS complexes activate RAS, leading to RAF?CMEK?CERK signaling. AKT phosphorylates FOXO1 and SREBP1, while mTORC1 promotes translation; the pathway also stimulates GLUT4 translocation. Attenuation is mediated by PTPN1/PTPRF phosphatases and SOCS1/3. INSR knockout disrupts these cascades, providing a loss-of-function model for dissecting insulin action.
In the context of 769-P VHL-mutant ccRCC cells, INSR knockout provides a unique tool to investigate the intersection of insulin signaling and cancer metabolism. Removal of insulin receptor input may unmask compensatory mechanisms involving AMPK or residual growth factor signaling and can help delineate the role of insulin in sustaining the aberrant metabolic phenotype of ccRCC. This model is especially valuable for exploring context-dependent oncogenic functions of insulin signaling and for testing combination therapies targeting both VHL-driven and insulin-mediated pro-tumorigenic pathways.
The knockout cells are suitable for investigating insulin resistance, metabolic reprogramming in ccRCC, and screening insulin-sensitizing agents. Use robust assays: western blotting for phospho-AKT and phospho-ERK, insulin-stimulated glucose uptake, RT-qPCR for metabolic genes, and metabolomics. For more information, contact Ascent Research.
