The IDH2 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the A-549 human lung adenocarcinoma cell line, featuring targeted disruption of the IDH2 gene. This polyclonal product comprises a heterogeneous pool of cells with CRISPR/Cas9-mediated gene disruptions, providing a robust loss-of-function model for studying isocitrate dehydrogenase 2 (IDH2) without clonal selection. The polyclonal format preserves biological variability for population-level metabolic and epigenetic analyses.
The A-549 parental cell line (ATCC CCL-185) is a human lung adenocarcinoma line with adherent epithelial morphology, widely used as a model for respiratory epithelium and non-small cell lung cancer. These cells display alveolar Type II pneumocyte characteristics and support studies of cancer metabolism, drug response, and oxidative stress in the lung epithelial context.
IDH2 encodes mitochondrial isocitrate dehydrogenase that catalyzes oxidative decarboxylation of isocitrate to ??-ketoglutarate, generating NADPH. This reaction sustains the TCA cycle, glutamine metabolism, and redox homeostasis by providing NADPH for reductive biosynthesis and glutathione-dependent antioxidant defense. IDH2 activity is regulated by HIF-1??, the deacetylase SIRT3, and cellular oxidative stress signals. The product ??-ketoglutarate is a co-substrate for TET dioxygenases and other ??-ketoglutarate-dependent enzymes, linking IDH2 to DNA demethylation. IDH2 interacts with IDH1, citrate synthase, aconitase, and mitochondrial chaperones HSP60/HSP10. Knockout of IDH2 disrupts ??-ketoglutarate synthesis, reduces NADPH availability, and perturbs downstream targets like TET2 and DNA methyltransferases, potentially altering the epigenome.
In the A-549 lung adenocarcinoma background, IDH2 knockout provides a system to study the intersection of central carbon metabolism, redox balance, and epigenetic regulation. Loss of IDH2 disrupts TCA flux and NADPH production, sensitizes cells to oxidative stress, and impairs ??-ketoglutarate-dependent dioxygenase activities, including those affecting DNA and histone methylation. This model is relevant for probing metabolic vulnerabilities, evaluating IDH2 as a drug target, and understanding mitochondrial contribution to tumorigenic phenotypes in lung epithelia.
This polyclonal knockout cell product is suited for cancer metabolism research, IDH2 inhibitor testing, epigenetic investigations, and oxidative stress studies. IDH2 disruption can be confirmed by western blot and RT-qPCR; metabolic reprogramming assessed via Seahorse flux analysis and LC-MS metabolite profiling (isocitrate, ??-ketoglutarate, citrate, succinyl-CoA); and functional phenotypes examined through NADPH/NADP+ ratio measurements, cell viability under oxidative challenge, colony formation, and proliferation assays. The polyclonal population serves as a reliable tool for drug target validation and pathway analysis. For further information, please contact Ascent Research.
