GPT2 Knockout A-549 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout population derived from the A-549 human lung adenocarcinoma epithelial cell line. This product introduces targeted disruption of the GPT2 gene encoding mitochondrial alanine aminotransferase, thereby eliminating GPT2 protein expression across a heterogeneous cell mixture. The polyclonal format provides a relevant model for population-level functional studies, avoiding biases inherent to clonal selection.
The parental A-549 cell line was originally established from the lung adenocarcinoma tissue of a 58-year-old Caucasian male and exhibits an epithelial morphology. A-549 cells are extensively used in cancer biology and respiratory research, serving as a model for lung adenocarcinoma signaling, drug metabolism, and epithelial barrier function. Their well-characterized metabolic profile and robust growth make them an excellent host for studying metabolic enzyme functions and adaptations.
GPT2 encodes mitochondrial alanine aminotransferase, a pyridoxal phosphate-dependent enzyme catalyzing the reversible conversion of alanine and ??-ketoglutarate to pyruvate and glutamate. This reaction is a critical link between amino acid catabolism and central carbon metabolism, providing anaplerotic substrates for the TCA cycle and gluconeogenic precursors. GPT2 expression is regulated by metabolic transcriptional regulators including PPARGC1A (PGC-1??), NRF1, PPAR??, and the glucocorticoid receptor NR3C1, and is responsive to mTOR and AMPK signaling pathways. The enzyme operates in the mitochondrial matrix, where it interacts with glutamate dehydrogenase (GLUD1/GLUD2) and TCA cycle enzymes, and its products pyruvate and glutamate contribute to energy production and glutathione synthesis. GPT2 functions within a broader network involving GOT1, GOT2, ASS1, ASL, PCK1, FBP1, and G6PC, underscoring its integration into alanine, aspartate, and glutamate metabolism, gluconeogenesis, and nitrogen handling.
In A-549 lung adenocarcinoma cells, knockout of GPT2 disrupts alanine-driven anaplerosis and glutamate homeostasis, potentially impairing mitochondrial function and redox balance. As cancer cells often rely on glutamine and alanine metabolism for proliferation and survival, loss of GPT2 forces metabolic rewiring that may expose vulnerabilities in lung adenocarcinoma. This model is therefore valuable for dissecting how amino acid metabolism supports cancer cell adaptability, drug resistance, and growth under nutrient-limited conditions.
This GPT2 knockout model supports diverse research applications, including elucidation of alanine metabolism in cancer cell growth and survival, investigation of mitochondrial amino acid metabolism, and modeling of GPT2 deficiency-linked neurological disorders. Researchers can examine metabolic reprogramming and drug resistance in lung adenocarcinoma, assess TCA cycle anaplerosis and redox alterations, and evaluate the impact on gluconeogenic pathways. Commonly used assays encompass Western blotting for GPT2, alanine transaminase activity measurements, LC-MS-based quantification of pyruvate and glutamate, targeted metabolomics of TCA intermediates, cell proliferation and apoptosis analysis, Seahorse respirometry for mitochondrial and glycolytic function, and RT-qPCR for metabolic gene expression. For additional technical information, please contact Ascent Research.
