The KYAT3 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from HEK293T, engineered to disrupt the KYAT3 gene. KYAT3 encodes a pyridoxal phosphate-dependent aminotransferase essential for kynurenine pathway flux. This knockout model eliminates endogenous KYAT3 activity, providing a genetically heterogeneous system that preserves the parental line??s high transfection efficiency and robust protein expression. The cells are supplied as a live stock, ready for expansion and use in a range of biochemical and cell-based assays.
The HEK293T parental line stably expresses SV40 large T antigen, enabling high-level transient protein expression and efficient virus production. Derived from human embryonic kidney HEK293 cells, this host is valued for its ease of culture, rapid growth, and lipid-based or calcium phosphate transfection compatibility. Large T antigen also supports episomal plasmid amplification via the SV40 origin of replication. The KYAT3 knockout polyclonal cells retain these traits, offering a versatile platform for studies requiring kynurenine metabolic flux analysis.
KYAT3 (kynurenine aminotransferase III) catalyzes the irreversible transamination of kynurenine to kynurenic acid, a neuroactive metabolite. It is transcriptionally regulated by HNF4?? and requires pyridoxal phosphate as cofactor, with kynurenine and glutamine as primary substrates. Kynurenic acid antagonizes NMDA receptors, negatively modulates ??7 nicotinic receptors, and agonizes GPR35. KYAT3 functions downstream of IDO1/TDO2, linking tryptophan catabolism to glutamatergic, cholinergic, and immune signaling. Its cysteine S-conjugate beta-lyase activity further connects it to sulfur amino acid metabolism.
Within HEK293T cells, KYAT3 knockout enables dissection of cell-autonomous kynurenic acid functions, free from neuronal or tissue-specific influences. Loss of kynurenic acid synthesis allows precise analysis of its impact on intracellular signaling, redox balance, and aryl hydrocarbon receptor activation. The polyclonal nature minimizes clonal bias and better reflects biological variability, enhancing translational relevance. These cells are an ideal background for reconstitution experiments, mutational analysis, and substrate specificity profiling of KYAT3.
This knockout model is suited for kynurenine pathway studies, neuroprotective drug screening, and cancer immunometabolism research. Representative assays include Western blotting, RT-qPCR, LC-MS-based kynurenic acid quantification, aminotransferase activity assays, NMDA receptor calcium flux, and AhR reporter assays. It supports investigation of glutamatergic signaling, immune evasion via tryptophan catabolism, and neurodegenerative disorders like schizophrenia. For further information, pricing, or technical consultation, contact Ascent Research.
