Ratiometric NAD+ Sensors Reveal Subcellular NAD+ Modulators

• DOI: 10.1021/acssensors.2c02565
• Publication Date: 2023-03-18T02:46:56Z
• Authors: Liuqing Chen; Meiting Chen; Mupeng Luo; Yong Li; Bagen Liao; Min Hu; Qiuliyang Yu
• Keywords: Cell Nucleus; 0303 health sciences; 03 medical and health sciences; Cytosol; Energy Transfer; Humans; NAD; Mitochondria

AbstractMapping NAD+ dynamics in live cells and human are essential for translating NAD+ interventions into effective therapies. Yet genetically encoded NAD+ sensors with better specificity and pH-resistance are still needed for cost-effective monitoring of subcellular and clinical NAD+. We introduce multicolor, resonance energy transfer-based NAD+ sensors that cover nano- to milli-molar concentration ranges for clinical NAD+ measurement and subcellular NAD+ visualization. The sensors captured the blood NAD+ increase induced by NMN supplementation and revealed the distinct subcellular effects of NAD+ supplements and modulators. Then, the sensors enabled high-throughput screenings for mitochondrial and nucleic NAD+ modulators and identified α-GPC, a cognition-related metabolite, induces NAD+ redistribution from mitochondria to nucleus relative to the total adenine nucleotides, which was confirmed by NAD+ FRET microscopy.