The IDH2 Knockout MCF-7 Polyclonal Cells represent a CRISPR/Cas9-mediated loss-of-function cell pool designed to disrupt the expression of isocitrate dehydrogenase 2 (IDH2) in the MCF-7 human breast adenocarcinoma line. This polyclonal knockout cell population provides a genetically heterogeneous model for interrogating IDH2-dependent metabolic and redox processes. Using non-homologous end joining repair after Cas9-induced double-strand breaks, the IDH2 gene is disrupted across the cell pool, enabling robust assessment of IDH2 function without clonal selection artifacts. The product is supplied as a ready-to-use, proliferating pool that maintains the epithelial morphology and key signaling hallmarks of the parental line while allowing direct evaluation of IDH2 contribution to mitochondrial metabolism and stress adaptation.
The MCF-7 host cell line is an estrogen receptor-positive (ER+) breast adenocarcinoma model established from the pleural effusion of a 69-year-old Caucasian female with metastatic mammary carcinoma. These cells are widely characterized for their hormone-dependent growth, intact p53 status, and responsiveness to estrogenic stimuli, making them a cornerstone in breast cancer research. MCF-7 cells retain many features of luminal A subtype breast tumors, including expression of ER?? and progesterone receptor, and they are frequently employed in studies of endocrine therapy resistance, cell cycle regulation, and apoptosis. The line??s adherent growth and well-documented metabolic profile facilitate the integration of IDH2 knockout-derived phenotypes into established experimental frameworks.
IDH2 encodes the mitochondrial isoform of isocitrate dehydrogenase, which catalyzes the oxidative decarboxylation of isocitrate to ??-ketoglutarate (??-KG) with concomitant reduction of NADP+ to NADPH. This reaction is a key NADPH-producing step in the TCA cycle and is critical for maintaining mitochondrial redox balance and biosynthetic capacity. IDH2 activity is regulated by deacetylation via SIRT3 and can be modulated by upstream factors such as HIF-1??, FoxO3a, and PGC-1??. The ??-KG produced serves as a substrate for TET2 dioxygenase and participates in glutaminolysis and reductive carboxylation pathways. IDH2-derived NADPH fuels glutathione reductase and glutathione peroxidase systems, directly linking mitochondrial metabolism to antioxidant defense. Interacting partners include mitochondrial complex I and the deacetylase SIRT3. Knockout of IDH2 abolishes this NADPH source, destabilizing redox homeostasis and sensitizing cells to oxidative insults.
In the MCF-7 context, IDH2 disruption offers a physiologically relevant platform for dissecting the metabolic vulnerabilities of ER+ breast cancer. The loss of IDH2 forces reliance on alternative NADPH-producing pathways, such as the pentose phosphate pathway or malic enzyme, and may alter ??-KG-dependent dioxygenase activities, including TET-mediated DNA demethylation. This model enables investigation of how IDH2 deficiency impacts tumor cell survival under oxidative stress, response to chemotherapeutics, and metabolic reprogramming. Given that IDH2 mutations are commonly observed in acute myeloid leukemia, glioma, and chondrosarcoma, the knockout cells also provide a comparative system for studying mutation-associated oncometabolite production and its consequences in a breast cancer background.
Researchers can employ these IDH2 Knockout MCF-7 Polyclonal Cells in a variety of assays, including NADPH/NADP ratio measurements, LC-MS-based metabolomic profiling of ??-KG and TCA cycle intermediates, and cell viability assessments under hydrogen peroxide challenge to evaluate oxidative stress resistance. The model is compatible with Seahorse metabolic flux analysis to quantify mitochondrial respiration and glycolytic shifts, as well as with Western blotting and RT-qPCR for verifying target disruption and compensatory pathway activation. Apoptosis detection via Annexin V/PI staining can further delineate IDH2??s role in cell death regulation. These applications make the cells a versatile tool for cancer metabolism studies, redox biology research, and drug response screening. For further information, please contact Ascent Research.
