The Cyp1a1 Knockout MODE-K Cell Line is a CRISPR/Cas9-edited knockout cell line featuring targeted disruption of the mouse Cyp1a1 gene within the MODE-K intestinal epithelial background. This loss-of-function model is generated using CRISPR/Cas9 technology to eliminate CYP1A1 expression, enabling precise dissection of its role in xenobiotic metabolism and aryl hydrocarbon receptor (AhR) signaling. The cell line is supplied as a ready-to-use, genomically defined population, optimized for in vitro studies of intestinal biology, toxicology, and carcinogenesis.
MODE-K cells are a well-established murine small intestinal epithelial cell line derived from C57BL/6 mice. They retain key characteristics of the intestinal epithelium, including the capacity for nutrient absorption, maintenance of barrier integrity, and participation in immune sensing. This background provides a physiologically relevant system for investigating gut-specific metabolic processes and epithelial responses to dietary and environmental compounds, making it an ideal host for studying the functions of CYP1A1 in a polarized, barrier-forming cell type.
Cyp1a1 encodes cytochrome P450 1A1, a phase I enzyme central to the oxidative metabolism of polycyclic aromatic hydrocarbons (PAHs) and other xenobiotics. Its expression is tightly controlled by the AhR/ARNT transcription factor complex, which is activated by ligands such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), benzo[a]pyrene, and dietary indole-3-carbinol. Upon activation, CYP1A1 catalyzes the formation of reactive intermediates that can form DNA adducts and induce oxidative stress. The enzyme interacts with NADPH-cytochrome P450 reductase and cytochrome b5 to mediate electron transfer, and its catalytic activity is commonly measured via the ethoxyresorufin-O-deethylase (EROD) assay. Ablation of Cyp1a1 therefore disrupts AhR-driven metabolic activation, blocking downstream genotoxic and oxidative damage pathways.
In the MODE-K intestinal context, Cyp1a1 knockout prevents the generation of genotoxic metabolites from dietary and environmental PAHs, thereby reducing DNA damage and oxidative stress within the epithelium. This model allows researchers to isolate CYP1A1-specific contributions to chemical carcinogenesis, inflammatory bowel disease, and colon cancer, separate from other AhR target genes. It also facilitates study of AhR-mediated gene regulation in gut homeostasis, as CYP1A1 is a sensitive marker of AhR activation. Without Cyp1a1, the effects of AhR ligands on intestinal barrier function and immune signaling can be examined without confounding metabolic activation, enabling clearer interpretation of AhR-dependent processes.
Researchers can employ this cell line for toxicology screening of PAHs and other xenobiotics, assessing endpoints such as DNA damage via comet assay and reactive oxygen species (ROS) detection. It supports investigation of AhR signaling through RT-qPCR of Cyp1a1 and AhR target genes, Western blotting for CYP1A1 protein, immunofluorescence staining, and barrier integrity assays. The model is valuable for studies on chemical carcinogenesis, inflammatory bowel disease, and colon cancer, providing a platform for evaluating intestinal metabolism of dietary xenobiotics in a controlled epithelial system. For additional information or technical support, please contact Ascent Research.
