KYNU Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with disrupted kynureninase (KYNU) gene expression in the human NCI-H1975 non-small cell lung adenocarcinoma cell line. This polyclonal format provides a heterogeneous pool of edited cells suitable for loss-of-function studies, avoiding assumptions of complete or clonal ablation of KYNU activity.
NCI-H1975 is an epithelial lung adenocarcinoma cell line harboring EGFR mutations L858R and T790M and a PIK3CA G118D missense mutation, while retaining wild-type TP53. These features make it a key model for EGFR-mutant NSCLC, particularly for investigating acquired resistance to tyrosine kinase inhibitors and the metabolic context of oncogenic signaling.
KYNU encodes a pyridoxal-5??-phosphate (PLP)-dependent kynureninase that hydrolyzes kynurenine and 3-hydroxykynurenine into anthranilic acid and 3-hydroxyanthranilic acid. This reaction is central to tryptophan catabolism, channeling metabolites toward NAD+ synthesis or complete oxidation. KYNU is regulated by inflammatory cytokines such as IFNG and TNF, and its activity influences downstream production of quinolinic acid and picolinic acid. KYNU operates within a network that includes IDO1, TDO2, KMO, HAAO, and QPRT, linking immune signals to cellular energy metabolism and AHR activation.
In NCI-H1975 cells, EGFR and PIK3CA mutations may intersect with KYNU-dependent metabolic reprogramming, potentially affecting NAD+ levels and the generation of immunosuppressive kynurenine derivatives. Disruption of KYNU permits dissection of how this enzyme contributes to metabolic plasticity and immune evasion, providing a relevant platform for studying the interplay between oncogenic drivers and tryptophan metabolism in lung adenocarcinoma.
Applications include LC-MS metabolomics for profiling kynurenine pathway intermediates, NAD+/NADH assays, cell proliferation analyses (MTT/CTG), as well as RT-qPCR and Western blotting for pathway enzyme expression. The polyclonal cells are also amenable to cytokine bead arrays, flow cytometry for immune markers, and xenograft tumor models. In combination with drug screens, this knockout model aids in exploring therapeutic strategies targeting NAD+ metabolism and tumor immunity. For additional product information, please contact Ascent Research.
