The IDH2 Knockout T-47D Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human breast ductal carcinoma epithelial cell line T-47D. This product features targeted disruption of the IDH2 gene, which encodes mitochondrial isocitrate dehydrogenase 2, using CRISPR/Cas9-mediated gene editing. The resulting polyclonal cells provide a heterogeneous loss-of-function model, enabling studies of IDH2-dependent metabolic pathways without the need for clonal isolation. This format is suitable for applications requiring bulk knockout populations, such as pooled screening and metabolic assays.
The parental T-47D cell line was established from a pleural effusion of a 54-year-old female with infiltrating ductal carcinoma of the breast. These cells are estrogen receptor-positive (ER+), progesterone receptor-positive (PR+), and express the luminal A molecular subtype markers, making them a key model for hormone-responsive breast cancer. They retain functional hormone receptor signaling and are widely used to study endocrine therapy resistance, tumor metabolism, and cell cycle regulation in an ER+ context.
IDH2 functions in the mitochondrial matrix as a homodimeric enzyme that catalyzes the oxidative decarboxylation of isocitrate to ??-ketoglutarate, coupled with the reduction of NADP+ to NADPH. This reaction bridges the TCA cycle and redox homeostasis, with IDH2 expression activated by SREBP1 and the PI3K/AKT pathway, and modulated by HIF-1??. The product ??-ketoglutarate is a co-substrate for TET dioxygenases, and NADPH is essential for fatty acid synthase (FASN) and glutathione (GSH) metabolism. IDH2 interacts with citrate synthase and aconitase, and functions downstream of IDH3. Disruption ablates ??-ketoglutarate and NADPH production, perturbing TCA cycle intermediates like citrate and succinyl-CoA.
In T-47D ER+ breast cancer cells, IDH2 knockout impairs mitochondrial metabolism, reducing ??-ketoglutarate and NADPH pools. This compromises lipid biosynthesis via FASN and weakens antioxidant defenses, elevating reactive oxygen species (ROS). The loss of ??-ketoglutarate may affect TET-mediated epigenetic regulation. Given the reliance of ER+ breast cancer on lipid and redox homeostasis, this model exposes metabolic vulnerabilities, potentially sensitizing cells to metabolic stress and chemotherapy. It serves as a platform to study synthetic lethality and drug responses in hormone-dependent cancers.
The cells support applications in cancer metabolism, redox biology, and drug discovery. Key assays include Seahorse metabolic flux analysis, LC-MS metabolomics, ROS detection, cell proliferation, Western blotting, and enzymatic activity measurements. They are suitable for metabolic vulnerability screening, glutamine metabolism studies, and high-throughput drug sensitivity assays. For further information, contact Ascent Research.
