IDH2 Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of human 143B osteosarcoma cells designed to disrupt the IDH2 gene, which encodes the mitochondrial isocitrate dehydrogenase 2 enzyme. This loss-of-function model enables investigation of IDH2-dependent metabolic and epigenetic processes in a cancer-relevant cellular context. The polyclonal nature of the knockout pool provides a heterogeneous mixture of edited alleles, facilitating robust functional studies without single-cell cloning bias.
The 143B host cell line is a highly metastatic human osteosarcoma line derived from HOS (human osteosarcoma) cells, widely employed as a model for bone cancer research and metastasis studies. Its aggressive phenotype and well-characterized genetic background make it suitable for dissecting molecular mechanisms driving tumor progression and metastatic dissemination. The osteosarcoma origin of 143B cells offers a physiologically relevant environment for examining IDH2 function in bone malignancies.
IDH2 is a mitochondrial NADP+-dependent enzyme catalyzing oxidative decarboxylation of isocitrate to ??-ketoglutarate (??-KG) with NADPH production in the TCA cycle. Its regulation involves the NAD+/NADH ratio, HIF-1??, and SIRT3 deacetylation. IDH2-supplied ??-KG is essential for TET1/TET2 DNA hydroxylases and Jumonji-domain histone demethylases like KDM4C, while NADPH fuels fatty acid synthesis via FASN and maintenance of reduced glutathione through glutathione reductase. Cofactors include NADP+ and Mg2+; interactions with the TOMM complex ensure mitochondrial localization. Knockout abrogates ??-KG and NADPH generation, disrupting fatty acid synthesis, antioxidant defense, and dioxygenase-mediated epigenetic remodeling.
In the 143B osteosarcoma background, IDH2 ablation provides a powerful tool to dissect the intersection between metabolism and malignancy. Loss of IDH2 activity diminishes NADPH availability, sensitizing cells to oxidative stress and potentially altering metastatic behavior. Decreased ??-KG levels can inhibit TET2-mediated DNA demethylation and KDM4C-mediated histone demethylation, leading to hypermethylation patterns that silence tumor suppressor genes. This model thus captures the consequences of IDH2 deficiency on TCA cycle flux, redox balance, and the epigenetic landscape, all within a bone cancer context.
This product is suitable for metabolic profiling of ??-KG, NADPH/NADP+, and 2-hydroxyglutarate, enzyme activity assays, and omics approaches like RNA-seq or metabolomics. Cell-based assays can assess proliferation, ROS levels, and apoptosis by flow cytometry. ChIP-qPCR enables evaluation of histone methylation changes, while Western blotting and RT-qPCR confirm gene expression. Applications include cancer metabolism, IDH2 drug validation, epigenetics, and redox studies. For support, contact Ascent Research.
