The IDH2 Knockout UM-UC-3 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the UM-UC-3 human bladder carcinoma cell line. This product enables targeted disruption of the IDH2 gene, which encodes mitochondrial isocitrate dehydrogenase 2, generating a heterogeneous pool of edited cells suitable for pooled loss-of-function studies. The polyclonal format retains cellular diversity while ensuring efficient target-gene knockout, facilitating robust experimental comparisons without clonal selection bias.
UM-UC-3 is a well-characterized transitional cell carcinoma line originally isolated from a male patient with bladder cancer. As an in vitro model for urothelial carcinoma, these cells exhibit properties consistent with aggressive cancer phenotypes, including anchorage-independent growth and invasive potential. The UM-UC-3 line has been extensively utilized to explore molecular mechanisms underlying bladder tumorigenesis, metastasis, and therapeutic resistance, making it a valuable platform for interrogating gene function via CRISPR/Cas9-mediated knockout.
IDH2 catalyzes the oxidative decarboxylation of isocitrate to ??-ketoglutarate, concomitant with NADP+ reduction to NADPH. This reaction is allosterically activated by ADP and regulated by deacetylase SIRT3, while transcription is promoted by FOXO3a. IDH2-derived NADPH serves as a critical reducing equivalent for glutathione reductase and thioredoxin reductase, maintaining cellular redox homeostasis and protecting against oxidative stress. Additionally, ??-ketoglutarate produced by IDH2 functions as a substrate for dioxygenases such as TET2 and prolyl hydroxylases, linking mitochondrial metabolism to epigenetic regulation and hypoxic signaling. IDH2 interacts with mitochondrial TCA cycle enzyme complexes and GRP75, positioning it at the intersection of energy metabolism and antioxidant defense.
In bladder cancer, metabolic reprogramming often involves altered IDH2 activity to support rapid proliferation and redox balance. Loss of IDH2 in UM-UC-3 cells disrupts mitochondrial NADPH generation, leading to compromised antioxidant capacity, altered TCA cycle flux, and potential reliance on compensatory pathways such as glutamine metabolism. This knockout model thus provides a physiologically relevant system to dissect IDH2-dependent metabolic vulnerabilities in urothelial carcinoma, with implications for targeting redox homeostasis in therapy-resistant tumors.
Researchers can employ these polyclonal knockout cells to investigate IDH2 function in bladder cancer metabolism, perform metabolic flux analyses using LC-MS metabolomics, assess redox status via glutathione and ROS assays, or evaluate mitochondrial respiration through Seahorse analysis. The model is suitable for drug target validation, particularly for agents that exploit redox imbalance or TCA cycle defects. Additional applications include studying IDH2 mutations relevant to glioma and acute myeloid leukemia by comparing bladder cancer metabolic adaptations. For further details regarding validation data or technical support, please contact Ascent Research.
