The KYNU Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited population of the human SK-HEP-1 liver adenocarcinoma cell line, featuring targeted disruption of the kynureninase (KYNU) gene. As a polyclonal product, it provides a heterogeneous mix of knockout alleles, offering a robust model for studying loss-of-function effects without clonal selection biases. This product is designed for advanced research on tryptophan catabolism and its role in hepatocellular carcinoma.
SK-HEP-1 cells, derived from ascitic fluid of a liver adenocarcinoma patient, exhibit epithelial morphology and high invasive potential, making them a widely used model for hepatocellular carcinoma metastasis and drug response studies. This cell line retains aggressive cancer hallmarks, enabling investigation of metabolic pathways that drive tumor progression.
KYNU encodes a PLP-dependent enzyme hydrolyzing kynurenine and 3-hydroxykynurenine to produce anthranilic and 3-hydroxyanthranilic acids, key intermediates in NAD+ biosynthesis. The enzyme operates downstream of IDO1 and TDO2 and interacts with KMO and QPRT. KYNU expression is regulated by IFN-??, TNF-??, and IL-1?? via STAT1, and its activity impacts AhR signaling, promoting transcription of CYP1A1, IL-22, and PD-1. Through these connections, KYNU links tryptophan metabolism to immune response modulation.
In SK-HEP-1 cells, KYNU disruption reduces 3-hydroxyanthranilic acid and NAD+ production while elevating kynurenine levels, which can hyperactivate AhR signaling. This alteration fosters an immunosuppressive microenvironment, enhancing expression of immune checkpoint molecules like PD-1. The polyclonal knockout population mirrors the heterogeneity of tumor cell populations, making it a relevant model to study metabolic immune evasion in hepatocellular carcinoma.
These cells enable detailed analyses of tryptophan metabolism and AhR signaling using LC-MS/MS metabolite quantification, luciferase reporter assays, NAD+/NADH measurements, and RT-qPCR for downstream targets (CYP1A1, IL-22). Co-culture with T cells and flow cytometry for PD-1 facilitate immune checkpoint research, while migration/invasion and metabolomics extend applications to cancer biology. For technical inquiries, contact Ascent Research.
