The IDH2 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the IDH2 gene in the human colorectal carcinoma cell line HCT 116. This polyclonal format preserves the cell line’s inherent genetic heterogeneity, providing a robust and reproducible loss-of-function model that avoids potential clonal bias. It is an ideal tool for studying the consequences of mitochondrial isocitrate dehydrogenase 2 ablation on cellular metabolism and redox regulation, making it suitable for a broad spectrum of metabolic and oncological investigations.
The HCT 116 host cell line is derived from a colorectal carcinoma and harbors KRAS G13D and PIK3CA oncogenic mutations, along with microsatellite instability (MSI-H) and mismatch repair (MMR) deficiency. These genetic features drive uncontrolled proliferation and create a dependency on metabolic reprogramming to sustain redox balance and biosynthesis, providing a medically relevant context for studying the impact of IDH2 knockout on tumor cell physiology.
IDH2 encodes a mitochondrial enzyme that catalyzes the oxidative decarboxylation of isocitrate to ??-ketoglutarate (??-KG), coupled with NADPH production. Its activity is regulated by upstream factors including HIF-1??, c-Myc, SREBP1, and SIRT3, and it cooperates with IDH1, citrate synthase, aconitase, and glutamate dehydrogenase. IDH2-derived NADPH is essential for maintaining glutathione (GSH) levels and controlling cellular reactive oxygen species (ROS); ??-KG serves as a co-substrate for dioxygenases that modulate histone methylation. Disruption of IDH2 compromises NADPH homeostasis, elevates ROS, and alters ??-KG-dependent epigenetic and metabolic processes.
In the HCT 116 background, IDH2 knockout disrupts mitochondrial TCA cycle function and depletes a key NADPH source, thereby increasing oxidative stress and impairing cellular proliferation. The sensitivity of these cells to redox perturbations, driven by oncogenic KRAS and PI3K signaling, makes this model particularly valuable for interrogating synthetic lethal interactions and metabolic vulnerabilities. The polyclonal composition enables population-level analyses of adaptive metabolic responses without the selectivity of single-cell clones.
These polyclonal knockout cells are compatible with diverse experimental techniques, including western blotting, RT-qPCR, NADPH/NADP+ ratio measurements, intracellular ROS detection, cell proliferation and clonogenic assays, apoptosis assays, metabolomics, and Seahorse metabolic flux analysis. They are well-suited for research in cancer metabolism, redox biology, drug sensitivity screening, metabolic reprogramming, and synthetic lethality, as well as for exploring IDH-related pathologies such as acute myeloid leukemia and glioma. For further details, please contact Ascent Research.
