INSR Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from HeLa cells, designed for targeted disruption of the insulin receptor gene. The heterogeneous pool contains diverse loss-of-function edits at the INSR locus, resulting in ablation of INSR protein expression. The polyclonal format avoids clonal selection bias and ensures broad mutation representation, supporting robust functional studies. These cells enable dissection of insulin receptor signaling in a human cervical adenocarcinoma context.
The HeLa cell line is an immortalized human cervical adenocarcinoma epithelial model, positive for human papillomavirus 18 (HPV-18) sequences. Widely used in cell biology and cancer research, HeLa cells offer robust growth and well-characterized signaling networks. HPV oncoproteins E6 and E7 degrade p53 and Rb, respectively, contributing to immortalization and providing a relevant background for oncogenic signaling studies. This parental line ensures compatibility with extensive experimental protocols and literature.
The INSR receptor tyrosine kinase mediates insulin action, with low-affinity binding to IGF1 and IGF2. Ligand-induced autophosphorylation activates adaptors IRS1, IRS2, and SHC, triggering the PI3K-Akt and MAPK/ERK cascades. The INSR??IRS1??PI3K??Akt pathway promotes glucose uptake and glycogen synthesis via GSK3?? inhibition, while INSR??SHC??Grb2??Ras??Raf??MEK??ERK drives proliferation. INSR interactions with Cbl and CAP facilitate receptor internalization and signal modulation. These pathways collectively regulate metabolism, growth, and survival.
INSR knockout in HeLa cells abolishes insulin-dependent Akt and ERK activation, impairing glucose uptake and altering proliferation. This model aids in dissecting INSR’s role in the metabolic reprogramming of cervical adenocarcinoma cells. Loss of mTOR and FOXO1 regulation affects protein synthesis and stress responses. The HeLa background, with active HPV oncogenes, allows examination of crosstalk between insulin and viral oncogenic pathways, as both converge on Akt and ERK.
Applications include mechanistic studies of insulin resistance, metabolic flux analysis, and inhibitor screening. Western blotting confirms INSR loss and monitors phospho-Akt, ERK, and GSK3??. Glucose uptake and proliferation assays quantify metabolic and growth changes. RNA-seq reveals transcriptomic alterations, while rescue experiments with PI3K or MEK inhibitors map pathway dependencies. The model supports research on chemotherapy resistance and PI3K-Akt-mTOR targeted therapies. For details, contact Ascent Research.
