INSR Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the HT29 human colorectal adenocarcinoma line, engineered to disrupt the INSR gene and eliminate insulin receptor expression. This polyclonal knockout model provides a heterogeneous loss-of-function system for studying insulin/IGF signaling in a cancer-relevant epithelial context.
The HT29 cell line, established from a stage III colorectal adenocarcinoma, is a widely used model of human intestinal epithelium and colorectal cancer. These adherent epithelial cells are tumorigenic and exhibit characteristic features of colorectal cancer, making them suitable for investigating cancer biology, drug response, and metabolic regulation.
The INSR gene encodes the insulin receptor, a receptor tyrosine kinase that mediates responses to insulin, IGF1, and IGF2. Upon ligand binding, the receptor autophosphorylates and recruits IRS1, IRS2, and SHC, activating the PI3K-AKT and MAPK/ERK (MAPK1/3) pathways. AKT phosphorylates downstream effectors including GSK3??, FoxO1, and mTORC1, thereby regulating glycogen synthesis, gluconeogenesis, and protein translation. PI3K activity promotes GLUT4 translocation to facilitate glucose uptake, while SREBP1c mediates lipogenic gene expression. The receptor also interacts with GRB2, PTP1B, SHP2, and caveolin1, integrating metabolic and mitogenic signals. INSR gene disruption abolishes these signaling cascades, eliminating insulin-stimulated glucose uptake, metabolic regulation, and proliferative signaling.
In the HT29 epithelial model, which harbors APC and TP53 alterations, INSR knockout removes key insulin-dependent inputs, enabling investigation of alternative growth and metabolic adaptation mechanisms. This system is particularly useful for studying metabolic reprogramming in colorectal cancer, including shifts in glucose and lipid utilization, and for assessing how tumors overcome insulin resistance to sustain proliferation. Furthermore, it provides a platform to explore resistance mechanisms to PI3K/AKT inhibitors by eliminating a major upstream activator.
Typical experimental approaches include western blot analysis of INSR, phospho-AKT (Ser473), and phospho-ERK (Thr202/Tyr204); fluorescent glucose uptake assays; RT-qPCR for GLUT4, SREBP1c, and FoxO1; MTT-based proliferation assays; colony formation; and xenograft tumor growth in nude mice. Insulin stimulation experiments confirm functional knockout, while metabolic flux analysis can reveal shifts in glycolytic and oxidative metabolism. These INSR Knockout HT29 Polyclonal Cells support research in cancer metabolism, drug resistance, and signal transduction. For technical details or ordering, please contact Ascent Research.
