The IDH1 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the human NCI-H1975 non-small cell lung adenocarcinoma cell line. Using CRISPR/Cas9-mediated gene disruption, the wild-type IDH1 gene is targeted to create a loss-of-function model without clonal isolation, providing a heterogeneous pool that reflects the genetic diversity of the edited population. This polyclonal format avoids potential clonal artifacts and enables robust assessment of IDH1-dependent cellular phenotypes in a physiologically relevant host background.
The NCI-H1975 cell line originates from the pleural effusion of a non-smoking female patient with lung adenocarcinoma and harbors activating EGFR L858R and resistance-conferring T790M mutations, making it a widely used model for studying EGFR tyrosine kinase inhibitor resistance. These cells maintain characteristic features of NSCLC and are suitable for investigating oncogenic signaling crosstalk with metabolic pathways. The integration of IDH1 knockout into this genetically defined NSCLC background allows examination of metabolic regulation in the context of EGFR-driven tumorigenesis.
IDH1 catalyzes the NADP+-dependent oxidative decarboxylation of isocitrate to ??-ketoglutarate, generating NADPH essential for reductive biosynthesis and antioxidant defense via glutathione metabolism. The enzyme also provides ??-ketoglutarate as a substrate for a broad family of dioxygenases, including prolyl hydroxylases that regulate HIF1A stability and Jumonji-domain histone demethylases that modulate chromatin modifications. IDH1 is regulated upstream by transcription factors such as HIF1A and Myc, as well as by substrate availability and post-translational modifications, while its activity feeds into pathways involving IDH2 and the ??-ketoglutarate dehydrogenase complex. Knockout of IDH1 ablates wild-type enzymatic function, leading to diminished NADPH pools and reduced ??-ketoglutarate availability, which in turn impairs cellular redox balance and alters dioxygenase-dependent processes affecting HIF signaling and epigenetic landscape.
In the NCI-H1975 background, loss of IDH1 creates a metabolic vulnerability that intersects with EGFR signaling. The reliance of lung cancer cells on NADPH for managing oxidative stress is particularly relevant under the selective pressure of drug-resistant phenotypes, where redox homeostasis is often rewired. This knockout model enables dissection of how IDH1-mediated NADPH production and ??-ketoglutarate-dependent dioxygenase activity contribute to cell survival, proliferation, and sensitivity to therapies. By perturbing these pathways, researchers can explore synthetic lethal interactions and metabolic dependencies unique to T790M-positive NSCLC.
The IDH1 Knockout NCI-H1975 Polyclonal Cells are suitable for a range of research applications, including investigations into redox metabolism, metabolic vulnerabilities in drug-resistant cancers, and the preclinical evaluation of IDH1-targeted therapies. Representative experimental approaches include NADPH/NADP+ quantification, ROS detection by flow cytometry, metabolomic profiling, Seahorse-based metabolic flux analysis, RT-qPCR, RNA-seq, western blotting, and cell viability assays under oxidative stress. Clonogenic assays provide functional readouts of long-term growth effects. For further technical details or to inquire about custom cell products, please contact Ascent Research.
