The INSR Knockout DLD-1 Polyclonal Cells are a CRISPR/Cas9-mediated polyclonal knockout population in the human DLD-1 colorectal adenocarcinoma cell line, with disruption of the insulin receptor gene (INSR). This loss-of-function model enables the study of insulin signaling in colon cancer. The polyclonal format maintains genetic diversity while eliminating INSR expression, making it suitable for bulk biochemical assays and pooled functional screens.
The parental DLD-1 line is a widely used colorectal adenocarcinoma model originally derived from a male patient, exhibiting epithelial morphology and bearing relevant oncogenic mutations, including APC truncation and KRAS G13D. It serves as a well-characterized host for cancer cell proliferation, apoptosis, and drug response studies, and its adherent growth facilitates robust CRISPR-based genetic manipulation.
INSR encodes a receptor tyrosine kinase activated by insulin, IGF1, and IGF2. Upon autophosphorylation, the receptor recruits insulin receptor substrates IRS1 and IRS2, triggering the PI3K-Akt cascade that regulates mTOR, FOXO1, and GLUT4-mediated glucose uptake. Parallel signaling through SHC1, GRB2, SOS1, RAS, RAF, MEK, and ERK promotes mitogenesis. Key negative regulators include PTP1B, SOCS1, and SOCS3, while adaptors such as GAB1 and PTEN further modulate signal output, placing INSR at a critical intersection of metabolic and growth control.
In DLD-1 colorectal cancer cells, INSR knockout disrupts insulin-dependent metabolic and mitogenic signaling, offering a platform to dissect insulin-specific functions in a KRAS-mutant context. This model enables exploration of crosstalk between oncogenic RAS and receptor tyrosine kinases, potential impacts on glucose metabolism, proliferation, and chemoresistance, and is relevant to research on insulin resistance and type 2 diabetes.
Representative experimental applications include Western blotting for phospho-Akt (Ser473) and phospho-ERK, glucose uptake assays using 2-NBDG, MTT viability tests, and apoptosis analysis by Annexin V/PI flow cytometry. Co-immunoprecipitation of INSR with IRS1, RT-qPCR for downstream targets such as GLUT4 and FOXO1, and metabolic profiling further characterize the knockout phenotype. These cells are well suited for drug screening efforts targeting insulin/IGF1R pathways. For ordering, technical support, or custom inquiries, please contact Ascent Research.
