IDH2 Knockout KYSE-30 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human esophageal squamous cell carcinoma line KYSE-30. This product provides targeted disruption of the IDH2 gene, which encodes mitochondrial isocitrate dehydrogenase 2, using a CRISPR/Cas9-based approach. The polyclonal population consists of a heterogeneous mix of edited cells carrying diverse loss-of-function mutations, offering a robust and unbiased model to study IDH2-dependent processes without clonal selection bias.
The KYSE-30 cell line originates from a well-differentiated human esophageal squamous cell carcinoma and exhibits epithelial morphology. It harbors a characterized TP53 mutation, a common feature in esophageal cancers that impairs genome stability and apoptosis. KYSE-30 is extensively used to investigate molecular mechanisms of esophageal squamous cell carcinoma, including proliferation, invasion, metabolism, and drug response.
IDH2 catalyzes the oxidative decarboxylation of isocitrate to ??-ketoglutarate (??-KG) in the mitochondrial matrix, generating NADPH. This TCA cycle reaction is critical for redox homeostasis, antioxidant defense, and reductive biosynthesis. IDH2 activity is modulated by the NAD+/NADH ratio and SIRT3 deacetylation. Downstream products ??-KG and NADPH are essential for ??-KG-dependent dioxygenases like TET2, linking IDH2 to DNA demethylation. IDH2 interacts with mitochondrial enzymes citrate synthase, aconitase, and glutamate dehydrogenase. Disruption of IDH2 reduces ??-KG and NADPH, impairing redox balance and dioxygenase function.
In esophageal squamous cell carcinoma, IDH2 supports the elevated metabolic and redox demands of cancer cells. IDH2 knockout in KYSE-30 depletes ??-KG, likely attenuating TET2-mediated DNA demethylation and altering genomic methylation patterns. Reduced NADPH compromises glutathione reduction, increasing susceptibility to oxidative stress. This model allows dissection of how IDH2 loss affects tumor cell viability, metabolic reprogramming, and sensitivity to redox-targeting chemotherapeutics.
This polyclonal knockout population enables detailed functional studies in esophageal cancer biology. Applications include quantifying NADPH and glutathione redox state, probing TET2 activity and DNA methylation profiles, and performing metabolomic and extracellular flux analyses to characterize metabolic reprogramming. The model also supports synthetic lethal screens and drug sensitivity testing with ROS-inducing or glutathione-depleting agents. Typical validation and readout methods include Western blotting, RT-qPCR, cell proliferation and apoptosis assays, and ROS detection. For further details or custom applications, contact Ascent Research.
