GOT1 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the GOT1 gene. The polyclonal pool provides a heterogeneous loss-of-function model that averages clonal variation, suitable for robust phenotypic screening and metabolic studies.
The NCI-H1975 host line is a human lung adenocarcinoma epithelial cell model derived from a female patient with non-small cell lung cancer, harboring the EGFR L858R/T790M double mutation. This clinically relevant background models acquired resistance to first-generation EGFR tyrosine kinase inhibitors and retains key oncogenic signaling, making it a valuable system for investigating metabolic adaptations in TKI-resistant NSCLC.
GOT1 codes for cytosolic aspartate aminotransferase, catalyzing the reversible transamination of aspartate and ??-ketoglutarate to oxaloacetate and glutamate. This reaction operates within the malate-aspartate shuttle, partnering with malate dehydrogenase 1 (MDH1) and the mitochondrial carriers SLC25A11 and SLC25A12 to transfer NADH reducing equivalents. GOT1 expression is activated by c-MYC and HIF1?? downstream of KRAS and EGFR signaling, and it supports TCA cycle anaplerosis, amino acid metabolism, and redox balance. By maintaining the cytosolic NAD+/NADH ratio and providing aspartate for nucleotide synthesis, GOT1 links oncogenic signals to biosynthetic and antioxidant programs in proliferating cells.
In NCI-H1975 carcinoma cells, GOT1-driven metabolic flux is thought to be critical for balancing redox demands imposed by EGFR-driven growth. Dysregulation of the malate-aspartate shuttle via GOT1 knockout may impair NADH oxidation and aspartate supply, sensitizing cells to oxidative stress and nutrient limitation. This model thus enables dissection of metabolic vulnerabilities in EGFR T790M-positive lung adenocarcinoma, particularly regarding TKI resistance and amino acid dependency.
Typical applications include stable isotope tracing with 13C-glutamine or 13C-glucose to map metabolic rewiring, quantitative metabolomics to measure oxaloacetate, glutamate, and aspartate pools, and NAD+/NADH assays to assess redox status. Proliferation assays under varying glutamine or aspartate availability can reveal nutrient dependencies, while drug sensitivity screens with EGFR TKIs help explore synthetic lethal interactions. Routine validation of GOT1 disruption should be performed by western blot or RT-qPCR. For technical inquiries, please reach out to Ascent Research.
