Subcellular characterization of NAD+ biosynthesis in metastatic melanoma by using organelle-specific biosensors

NAD+ plays central roles in a wide array of normal and pathological conditions. Inhibition of NAD+ biosynthesis can be exploited therapeutically in cancer, including melanoma. To obtain quantitation of NAD+ levels in live cells and to address the issue of the compartmentalization of NAD+ biosynthesis, we exploited a recently described, genetically-encoded NAD+ biosensor (LigA-cpVENUS), which was targeted to the cytosol, mitochondria and nuclei of BRAF-V600E A375 melanoma cells, a model of metastatic melanoma (MM).