The BCAT2 Knockout A-549 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout population carrying a targeted disruption of the BCAT2 gene. This loss-of-function model eliminates branched-chain amino acid transaminase 2 expression, enabling investigation of BCAA metabolism and its downstream effects. The polyclonal format provides a genetically heterogeneous pool of null cells, suitable for bulk functional analyses without clonal bias.
The A-549 host line is an established human lung adenocarcinoma epithelial line derived from a 58-year-old male, characterized by a hypo-triploid karyotype. It serves as a widely used model for non-small cell lung adenocarcinoma (NSCLC), recapitulating key oncogenic signaling and metabolic features. A-549 cells are commonly employed to study lung cancer biology, therapeutic resistance, and metabolic reprogramming.
BCAT2 encodes a mitochondrial branched-chain amino acid transaminase that catalyzes the reversible transamination of leucine, isoleucine, and valine to their respective branched-chain keto acids, using pyridoxal phosphate (PLP) as a cofactor. The reaction provides carbon substrates for the TCA cycle and regulates mTORC1 activation via the mTORC1/S6K1 axis. BCAT2 expression is controlled by upstream regulators such as c-Myc, HIF-1??, and mTORC1, and its activity functionally interacts with the BCKDH complex. BCAT2-mediated BCAA catabolism influences acetyl-CoA levels and histone acetylation machinery, thereby linking metabolism to epigenetic regulation.
In A-549 NSCLC cells, BCAT2 plays a pivotal role in sustaining mitochondrial metabolism and nutrient-sensing pathways. Disruption of BCAT2 in these polyclonal cells impairs BCAA catabolism, reducing TCA cycle anaplerosis and potentially attenuating mTORC1 signaling. This model is therefore valuable for dissecting metabolic vulnerabilities in lung adenocarcinoma and for exploring the crosstalk between BCAA metabolism and oncogenic signaling networks driven by mTORC1 and c-Myc.
Researchers can utilize BCAT2 Knockout A-549 Polyclonal Cells for a variety of experiments, including BCAA quantification, TCA cycle metabolite profiling, and mTORC1 phospho-signaling analysis by western blotting for phospho-S6K1. Functional studies such as cell proliferation, migration, and drug sensitivity assays, combined with Seahorse metabolic flux analysis and glutamine/glutamate measurement, enable comprehensive characterization of metabolic reprogramming. The knockout model supports therapeutic target validation and metabolic inhibitor screening in the context of NSCLC. For further information or technical support, please contact Ascent Research.
