The IDH3G Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa human cervical adenocarcinoma cell line, engineered to disrupt the IDH3G gene. This product provides a mixed population of cells with heterogeneous gene-editing events at the IDH3G locus, enabling the study of IDH3G loss-of-function effects in a widely used epithelial cancer model. The knockout is achieved through CRISPR/Cas9-mediated gene disruption, resulting in a loss-of-function model suitable for bulk biochemical and functional assays.
The host cell line, HeLa, is an immortalized epithelial cell line originally isolated from an African American woman with cervical carcinoma, and it contains integrated HPV-18 genomic sequences. These cells express viral oncoproteins E6 and E7, which inactivate p53 and Rb, respectively, driving continuous proliferation. HeLa cells are extensively employed in cancer biology, virology, and metabolic research, offering a well-characterized background for investigating mitochondrial TCA cycle alterations and metabolic reprogramming in a tumorigenic context.
IDH3G encodes the gamma subunit of the mitochondrial NAD+-dependent isocitrate dehydrogenase 3 (IDH3) complex, which catalyzes the oxidative decarboxylation of isocitrate to alpha-ketoglutarate (??-KG) while reducing NAD+ to NADH. The IDH3 holoenzyme includes catalytic subunits IDH3A and IDH3B, with IDH3G acting as a regulatory subunit. Activity is allosterically activated by citrate, ADP, and calcium, and inhibited by a high NAD+/NADH ratio. Transcription is regulated by PGC-1??, NRF1, TFAM, and ERR??, linking IDH3G expression to mitochondrial biogenesis. NADH produced fuels ATP synthesis via the electron transport chain, and ??-KG serves as a substrate for ??-ketoglutarate-dependent dioxygenases such as TET and Jumonji demethylases, connecting TCA metabolism to epigenetic regulation.
In the HeLa cervical cancer background, which displays elevated glycolytic flux and mitochondrial dysfunction common in tumors, IDH3G knockout is expected to reduce TCA cycle flux, lowering ??-KG and NADH production and impairing oxidative phosphorylation. This metabolic perturbation can reveal vulnerabilities in cancer cells reliant on residual TCA cycle activity for biosynthesis, redox homeostasis, or epigenetic maintenance. Moreover, the model provides a tool to dissect how IDH3 dysfunction may contribute to diseases such as retinitis pigmentosa, where IDH3G mutations have been implicated, and to explore compensatory metabolic rewiring.
This polyclonal knockout product is suited for metabolomic profiling by LC-MS to quantify TCA cycle intermediates, mitochondrial respiration assays using Seahorse analyzers, and NAD+/NADH ratio measurements. Researchers can assess the impact of IDH3G loss on ??-KG-dependent dioxygenase activity, epigenetic marks, cancer cell proliferation, and apoptosis. Additional applications include synthetic lethal screening, evaluating metabolic dependencies, and performing transcriptomic analyses via RNA-seq. For further technical inquiries or custom applications, please contact Ascent Research.
