The IDH1 Knockout HeLa Cell Line is a CRISPR/Cas9-edited human cell line with targeted disruption of the IDH1 gene. This loss-of-function model is provided as a ready-to-use live cell line for investigating IDH1-dependent metabolic and signaling processes in a cervical carcinoma background.
HeLa is a widely used human epithelial cell line derived from a cervical adenocarcinoma of a 31-year-old African American female. The cells harbor integrated HPV-18 sequences, which contribute to their transformed phenotype, and serve as a robust model system for cancer research, particularly for studies on HPV-driven oncogenesis and signal transduction.
IDH1 encodes the cytosolic and peroxisomal isocitrate dehydrogenase 1, which catalyzes the oxidative decarboxylation of isocitrate to ??-ketoglutarate (??-KG) with concomitant reduction of NADP+ to NADPH. This reaction is integral to the citrate cycle, cellular energy metabolism, and maintenance of redox homeostasis. IDH1 activity is regulated by upstream factors including HIF-1??, c-Myc, and SREBP1, and its product ??-KG serves as a co-substrate for dioxygenases such as TET family DNA demethylases and histone lysine demethylases. Additionally, IDH1-generated NADPH supports glutathione metabolism and defense against oxidative stress. The enzyme interacts physically with its substrate isocitrate and cofactor NADP+, and functionally with the mitochondrial isoforms IDH2 and IDH3 to coordinate cellular metabolism.
Disruption of IDH1 in HeLa cells abolishes wild-type enzymatic function, leading to reduced ??-KG and NADPH pools. This perturbs redox balance, increases susceptibility to oxidative stress, and impairs the activity of ??-KG-dependent dioxygenases such as TET2 and histone demethylases, which may result in altered DNA and histone methylation patterns. Given that IDH1 mutations are frequent in gliomas, acute myeloid leukemia, chondrosarcoma, and related conditions, the knockout cell line provides a clean loss-of-function background to dissect IDH1??s tumor-suppressive or metabolic roles independent of dominant-negative oncometabolite production.
Researchers can employ this model to study cancer metabolism, epigenetic regulation, and the consequences of NADPH/??-KG imbalance. Representative assays include Western blotting and RT-qPCR for confirmatory gene expression analysis, ??-KG quantification, NADPH/NADP+ ratio measurements, cell proliferation, ROS detection, and DNA methylation profiling. The line is suitable for drug target validation and can be used to screen compounds that modulate metabolic vulnerabilities. For further information or to request this product, please contact Ascent Research.
