IDH3G Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population targeting the IDH3G gene in human SK-HEP-1 liver adenocarcinoma cells. This heterogeneous pool of cells carries diverse gene disruptions, allowing loss-of-function studies of the mitochondrial isocitrate dehydrogenase gamma subunit. The polyclonal format reduces clonal artifacts and is ideal for pooled screening and functional genomics. The cells are supplied validated at the population level and ready for TCA cycle and metabolism research.
The SK-HEP-1 cell line, derived from ascites of a liver adenocarcinoma patient, displays endothelial characteristics and serves as a model for liver sinusoidal endothelial cells (LSECs). It retains metabolic hallmarks of hepatic tissue, including oxidative phosphorylation and glycolysis, making it a robust system for studying mitochondrial function and metabolic reprogramming in liver diseases and cancer.
IDH3G encodes the gamma subunit of mitochondrial NAD+-dependent isocitrate dehydrogenase, which forms a heterotetramer with IDH3A and IDH3B. This complex catalyzes the irreversible decarboxylation of isocitrate to ??-ketoglutarate (??-KG) and generates NADH. IDH3G activity is regulated by hypoxia-inducible factors, nutrient status, and mitochondrial biogenesis signals. Downstream, ??-KG serves as a substrate for ??-KG-dependent dioxygenases and an anaplerotic precursor. Disruption of IDH3G impairs ??-KG and NADH production, reducing electron transport chain flux and altering redox balance. Its interaction with IDH3A and IDH3B is critical for complex stability and catalytic function.
In SK-HEP-1 cells, IDH3G knockout provides a model to study TCA cycle dysfunction in a liver cancer context. Loss of IDH3G diminishes ??-KG and NADH, potentially triggering metabolic adaptation and affecting ??-KG-dependent dioxygenase activities such as demethylation. This system can elucidate how mitochondrial impairment influences endothelial-like properties and liver sinusoidal biology, and offers a tool to investigate metabolic underpinnings of retinitis pigmentosa-linked mitochondrial deficits.
These polyclonal knockout cells are suitable for a range of experimental workflows, including immunoblotting and RT-qPCR to verify IDH3G loss, oxygen consumption rate assays to measure mitochondrial respiration, and mass spectrometry-based metabolomics to profile TCA cycle intermediates. Additional applications utilize cell viability and ROS detection to assess metabolic stress. Researchers can employ this model to investigate liver cancer metabolic vulnerabilities, ??-KG-dependent dioxygenase signaling, and mitochondrial dysfunction. For further information or customization, please contact Ascent Research.
