The IDH2 Knockout KYSE-150 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the IDH2 gene in the human KYSE-150 esophageal squamous cell carcinoma line. The polyclonal format provides a heterogeneous pool of loss-of-function variants, minimizing clonal selection bias and offering a robust model for studying mitochondrial enzyme function.
KYSE-150 is an epithelial cancer cell line derived from human esophageal squamous cell carcinoma, widely used in oncology to investigate tumor metabolism, drug sensitivity, and disease mechanisms. Esophageal squamous cell carcinoma cells often rely on mitochondrial activity and redox balance, making this background relevant for exploring IDH2??s roles in cancer metabolism. The cell line??s genetic stability and well-characterized phenotype support reproducible experimental outcomes.
IDH2 is a mitochondrial enzyme that catalyzes the NADP+-dependent oxidative decarboxylation of isocitrate to ??-ketoglutarate (??-KG) using Mg2+ or Mn2+ cofactors, producing NADPH essential for antioxidant defense and reductive biosynthesis. Its expression is regulated by transcription factors FOXO and HIF1?? and is responsive to the NAD+/NADH ratio. IDH2 functions within the citric acid cycle alongside aconitase and citrate synthase, while its product ??-KG is further metabolized by glutamate dehydrogenase, linking to glutamine metabolism. Mutations in IDH2 leading to 2-hydroxyglutarate accumulation are implicated in various cancers.
In KYSE-150 cells, disruption of IDH2 compromises mitochondrial NADPH generation, likely heightening vulnerability to oxidative stress and altering proliferative signaling. This knockout model enables dissection of IDH2-dependent contributions to cancer cell redox homeostasis, metabolic plasticity, and survival under stress. It also provides a platform to uncover compensatory pathways, such as glycolytic upregulation, and to test the dependency of esophageal carcinoma cells on IDH2 activity. The polyclonal population captures heterogeneous editing outcomes, reflecting tumor heterogeneity.
Applications include metabolic flux analysis (Seahorse) to assess oxygen consumption and acidification, NADPH/NADP+ ratio measurements, and ??-KG quantification. Researchers can perform western blotting, RT-qPCR, proliferation, apoptosis, and colony formation assays to evaluate functional consequences. Immunofluorescence can detect changes in mitochondrial physiology. The model is valuable for drug target validation, particularly IDH2 inhibitor testing, and for biomarker discovery related to redox metabolism. For further information, contact Ascent Research.
