The INSR Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the human INSR gene. This product delivers a heterogeneous pool of HEK293T cells with targeted disruption of the insulin receptor locus, avoiding the need for single-cell cloning while maintaining a functional knockout across the population. It is optimized for population-level assays in metabolic and signaling research.
HEK293T is a human embryonic kidney cell line expressing the SV40 large T antigen, derived from the parental HEK293 line. It is valued for its high transfection efficiency, rapid proliferation, and robust protein production capacity, making it a workhorse for recombinant protein expression and lentiviral packaging. Although not a classical insulin-responsive tissue, HEK293T cells express core insulin signaling components, enabling the dissection of insulin receptor function in a genetically tractable background.
INSR encodes the insulin receptor, a receptor tyrosine kinase that mediates insulin??s metabolic and mitogenic actions. Insulin binding triggers receptor autophosphorylation and activation of two principal pathways: the IRS1/PI3K/AKT cascade and the RAS/MAPK cascade. Key downstream effectors include IRS1, PI3K, AKT, GSK3, FOXO, mTOR, S6K, and the ERK1/2 kinases. The receptor also engages adaptors such as SHC1 and GRB2 to propagate signals. INSR knockout ablates these insulin-dependent signals, offering a null background for mechanistic studies.
In HEK293T cells, disruption of INSR eliminates endogenous insulin receptor activity without affecting viability, as these cells do not depend on insulin for growth. This loss-of-function model permits unambiguous assignment of signaling events to the insulin receptor, facilitates the study of insulin receptor mutants and biased agonists, and enables investigation of receptor crosstalk with other tyrosine kinases. The polyclonal design reduces clonal bias, making the product well-suited for screening campaigns and quantitative dose?Cresponse analyses.
Research applications include the investigation of insulin resistance, type 2 diabetes, and receptor tyrosine kinase signaling. Common assays are insulin-stimulated phospho-AKT western blot, glucose uptake assays, RT-qPCR of downstream targets (e.g., GLUT4, FOXO1), and co-immunoprecipitation of IRS1 or PTPN1. Further applications encompass immunofluorescence for receptor trafficking, phospho-signaling arrays, and flow cytometry for surface INSR detection. These cells are useful for validating anti-diabetic agents and modeling genetic insulin resistance. For more details, contact Ascent Research.
