INSR Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human INSR gene. This loss-of-function model is generated by CRISPR/Cas9-mediated gene disruption in HAP1 cells, eliminating expression of the insulin receptor. The polyclonal format provides a heterogeneous mixture of knockout alleles, suitable for population-based functional assays, pooled screens, and studies requiring robust representation of diverse editing outcomes.
The HAP1 host cell line is a near-haploid, adherent, fibroblast-like line derived from the male KBM-7 chronic myeloid leukemia background. Its haploid genome simplifies gene editing and facilitates the generation of reliable knockout models, making it a cornerstone in functional genomics and genetic screening. HAP1 cells retain key signaling pathways, allowing relevant investigation of metabolic and growth-regulating mechanisms within a genetically tractable system.
The INSR gene product, insulin receptor, is a receptor tyrosine kinase that plays a central role in metabolic control. Ligands such as insulin, IGF1, and IGF2 induce receptor autophosphorylation, recruiting key adaptors including IRS1 and SHC1. Activated IRS1 couples the receptor to the PI3K-AKT pathway, leading to GLUT4 translocation and glucose uptake, while SHC1 links to the RAS-RAF-MEK-ERK cascade, promoting cell proliferation. Additional interacting factors like GRB2, SOS, and the phosphatase PTPN1 fine-tune signal strength. Downstream targets IRS2, AKT, and ERK execute metabolic and mitogenic programs, highlighting INSR??s pivotal role in balancing metabolic and growth responses.
In the context of HAP1 cells, knockout of INSR creates a null insulin-signaling environment, rendering the cells unresponsive to insulin stimulation. This ablation mimics severe insulin resistance and is instrumental for modeling pathologies such as diabetes mellitus, Rabson-Mendenhall syndrome, Donohue syndrome, and type A insulin resistance. The combination of INSR deficiency with the haploid background ensures unambiguous interpretation of insulin-dependent phenotypes, circumventing compensatory signaling often observed in diploid systems.
INSR Knockout HAP1 Polyclonal Cells are ideally suited for biochemical and functional analyses. Typical applications include Western blotting to monitor phosphorylation of IRS1 and AKT under insulin treatment, glucose uptake assays to quantify metabolic activity, and RT-qPCR to measure insulin-regulated gene expression. Additionally, phospho-proteomic workflows and metabolic flux analyses can dissect pathway activities. These applications support drug target validation, functional genomics, and mechanistic studies of insulin action. For further information on product specifications and experimental protocols, please contact Ascent Research.
