The KMO Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human colorectal carcinoma cell line HCT 116. This loss-of-function model disrupts the kynurenine 3-monooxygenase (KMO) gene, a central enzyme in the kynurenine pathway of tryptophan metabolism, enabling the study of KMO-dependent metabolic and signaling processes.
The HCT 116 host cell line is an epithelial colorectal carcinoma model featuring microsatellite instability, a KRAS G13D mutation, wild-type TP53, and a CTNNB1 mutation that stabilizes ??-catenin. These characteristics make it an established system for investigating oncogenic signaling, Wnt pathway activation, and metabolic dysregulation in colorectal cancer.
KMO is an FAD-dependent monooxygenase that converts kynurenine to 3-hydroxykynurenine, a critical step determining the balance of neuroactive and immunomodulatory metabolites. KMO expression is activated by interferon-gamma (IFNG), tumor necrosis factor (TNF), and STAT1, and is responsive to toll-like receptor agonists. Downstream metabolites include 3-hydroxykynurenine, quinolinic acid, and picolinic acid, whereas diminished KMO activity shunts kynurenine toward kynurenic acid, an NMDA receptor antagonist. The enzyme interacts with kynureninase and 3-hydroxyanthranilic acid dioxygenase and functions within a network that includes indoleamine 2,3-dioxygenase (IDO) and tryptophan 2,3-dioxygenase (TDO), ultimately regulating NAD+ biosynthesis.
Knockout of KMO in HCT 116 cells disrupts the kynurenine pathway, causing kynurenine accumulation and enhanced kynurenic acid production. This metabolic reprogramming can impact tumor cell proliferation, migration, and immune evasion by altering the local metabolite milieu. The model enables dissection of how KRAS and ??-catenin-driven signaling cooperates with tryptophan metabolism to shape tumor behavior.
These polyclonal knockout cells are suitable for metabolic profiling by LC-MS, drug sensitivity testing with KMO inhibitors, and flow cytometric analysis of immune checkpoint molecules. Additionally, they can be used in proliferation and migration assays, as well as RT-qPCR and western blotting to validate target disruption and downstream pathway effects. For further technical details, please contact Ascent Research.
