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Ratiometric NAD+ sensors reveal subcellular NAD+ modulators

Chen, L.; Chen, M.; Luo, M.; Li, Y.; Liao, B.; Hu, M.; Yu, Q.

2022-05-20 bioengineering
10.1101/2022.05.20.491061 bioRxiv
Show abstract

Mapping 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 -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.

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