The IDE Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of HAP1 cells featuring targeted disruption of the IDE gene. This loss-of-function model is designed for studying insulin-degrading enzyme (IDE) biology in a near-haploid human background. The polyclonal format provides a heterogeneous pool of IDE-disrupted cells, facilitating reproducible, population-level analyses without clonal selection. It is ideal for functional genomics assays and pathway validation.
The host HAP1 cell line is a near-haploid human cell line derived from chronic myelogenous leukemia KBM-7 cells. With a predominantly haploid karyotype, HAP1 cells are exceptionally suited for CRISPR-mediated gene knockout because a single targeting event can abolish gene function. They are widely used in genetic screens and knockout studies, supporting high-throughput formats and enabling unambiguous genotype-phenotype correlations.
IDE codes for a zinc metalloprotease that degrades insulin, glucagon, amyloid-beta, bradykinin, and atrial natriuretic peptide. Its expression is regulated by insulin, PPARG agonists (e.g., rosiglitazone), FOXO1, glucose, and HNF4A. IDE directly interacts with these substrates and the insulin receptor, influencing receptor recycling and signal termination. In insulin signaling, IDE operates downstream of INSR and upstream of AKT, while in Alzheimer??s?related pathways it contributes to amyloid-beta clearance alongside APP, BACE1, and PSEN1. Thus, IDE acts at a nexus between metabolic and neurodegenerative signaling.
Disrupting IDE in the haploid HAP1 background eliminates wild-type allele interference, allowing precise dissection of IDE??s role in peptide degradation. This model enables investigation of how loss of IDE function affects insulin clearance, receptor recycling, and amyloid-beta accumulation. The polyclonal nature mimics biological variability, making it relevant for studying insulin resistance and Alzheimer??s disease mechanisms in a clean genetic system.
Applications include insulin and amyloid-beta degradation assays, ELISA quantification, co-immunoprecipitation to probe IDE-substrate binding, and phospho-AKT analysis to assess insulin pathway activity. RT?qPCR and Sanger sequencing confirm gene disruption, while flow cytometry monitors insulin receptor expression. This product supports drug target validation, functional genomics screens, and metabolic disease research. For additional details, contact Ascent Research.
