IRS1 Knockout A-549 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population generated from the A-549 human lung adenocarcinoma epithelial line. The product is produced via CRISPR/Cas9-mediated gene disruption of the IRS1 locus, yielding a heterogeneous pool that collectively models loss of IRS1 function. This polyclonal format avoids clonal bias and enables robust population-level analyses of deficient insulin/IGF-1 signaling. It is suited for dissecting IRS1-dependent cancer cell biology, metabolic regulation, and signal transduction, offering a tool for pathway analysis and drug response profiling.
The parental A-549 cell line, derived from lung adenocarcinoma tissue of a 58-year-old Caucasian male, exhibits adherent epithelial morphology and alveolar type II pneumocyte characteristics. Widely used in cancer research, A-549 cells retain insulin and IGF-1 sensitivity, making them a relevant host for studying IRS1 in lung cancer proliferation, survival, and metabolism. This background provides a clinically relevant context for examining IRS1-dependent pathways in non-small cell lung cancer and associated metabolic disorders.
IRS1 encodes an adaptor protein functioning as a key docking molecule downstream of the insulin receptor (INSR) and IGF-1 receptor (IGF1R). Ligand-activated receptors phosphorylate IRS1, recruiting effectors including the p85 regulatory subunit of PI3K (PIK3R1) and GRB2, thereby activating the PI3K/AKT and RAS/RAF/MEK/ERK (MAPK1/3) cascades. AKT-mediated mTORC1 activation and FOXO1 phosphorylation regulate metabolism, while ERK1/2 promotes proliferation. IRS1 also interacts with SHP2 and 14-3-3 proteins to modulate signal duration. Inflammatory cytokines, such as TNF-alpha and IL-6, can inhibit IRS1 function, linking insulin resistance to cancer biology. Thus, IRS1 integrates metabolic and mitogenic signals.
In A-549 cells, IRS1 knockout disrupts canonical insulin/IGF-1 transduction, enabling dissection of IRS1-dependent proliferative, survival, and metabolic pathways. Loss of IRS1 impairs AKT1 and ERK1/2 activation, attenuating mTORC1 signaling and altering glucose metabolism, which highlights metabolic vulnerabilities in cancer. This model facilitates investigation of crosstalk between growth factor signaling and metabolic reprogramming in lung cancer, providing insights into resistance mechanisms and the contribution of insulin/IGF-1 pathways to tumor progression. It serves as a valuable tool for validating IRS1 as a molecular target in oncology and metabolic disease.
These knockout cells are applicable to western blotting for IRS1 and phospho-AKT, RT-qPCR for IRS1 mRNA, MTT or BrdU proliferation assays, glucose uptake measurements, and migration/invasion studies. Co-immunoprecipitation can assess disrupted IRS1-p85 interactions, while phospho-flow cytometry and drug sensitivity testing with insulin/IGF-1 inhibitors characterize pathway rewiring. They are ideal for metabolic regulation studies, drug response profiling, and research on diabetes-related cancer biology. For detailed specifications and support, please contact Ascent Research.
