The GOT1 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from A-549 human lung carcinoma cells, designed for loss-of-function analysis of GOT1. This polyclonal model provides genetically heterogeneous target-gene disruption, reducing clonal bias and better capturing population-level metabolic phenotypes in epithelial tumor research.
The A-549 cell line is an epithelial adenocarcinoma model from a 58-year-old male, extensively used to study lung cancer metabolism, metastasis, and therapeutic resistance. Its KRAS and TP53 mutations make it especially relevant for investigating how metabolic enzymes like GOT1 support tumorigenesis.
GOT1 encodes cytosolic aspartate aminotransferase, catalyzing reversible conversion of aspartate and ??-ketoglutarate to oxaloacetate and glutamate??a key step in the malate-aspartate shuttle. This process transfers reducing equivalents across mitochondria, linking glycolysis to gluconeogenesis and maintaining NAD+/NADH balance. GOT1 is regulated by NRF2, ATF4, c-MYC, and mTORC1, and it modulates downstream pools of aspartate, glutamate, and oxaloacetate while interacting with MDH1 and GOT2. These activities coordinate amino acid biosynthesis with central carbon flux, fueling nucleotide production and redox homeostasis in proliferating cells.
In A-549 cells, GOT1 disruption enables dissection of metabolic vulnerabilities arising from malate-aspartate shuttle dysfunction. Loss of GOT1 impairs cytosolic aspartate utilization, limiting nucleotide biosynthesis and potentially sensitizing cells to nutrient stress. Given the frequent hyperactivation of NRF2 and mTORC1 in non-small cell lung cancer, this knockout model allows systematic study of adaptive mechanisms in redox control and drug resistance, including pathways that circumvent aspartate depletion under chemotherapy.
Applications include metabolic flux analysis using stable isotopes, LC-MS amino acid profiling, Seahorse respirometry, and NAD+/NADH measurement to probe redox shifts. Standard validation by Western blot, RT-qPCR, co-IP with MDH1, and proliferation assays (MTT, BrdU) supports mechanistic studies. The product is valuable for anticancer drug screening, metabolic reprogramming research, and investigating tumor cell dependencies on amino acid metabolism. For additional information, contact Ascent Research.
