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. 2020 Apr 20;53(2):240-252.e7.
doi: 10.1016/j.devcel.2020.02.017. Epub 2020 Mar 19.

Illuminating NAD + Metabolism in Live Cells and In Vivo Using a Genetically Encoded Fluorescent Sensor

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Illuminating NAD + Metabolism in Live Cells and In Vivo Using a Genetically Encoded Fluorescent Sensor

Yejun Zou et al. Dev Cell. .

Abstract

Understanding of NAD+ metabolism provides many critical insights into health and diseases, yet highly sensitive and specific detection of NAD+ metabolism in live cells and in vivo remains difficult. Here, we present ratiometric, highly responsive genetically encoded fluorescent indicators, FiNad, for monitoring NAD+ dynamics in living cells and animals. FiNad sensors cover physiologically relevant NAD+ concentrations and sensitively respond to increases and decreases in NAD+. Utilizing FiNad, we performed a head-to-head comparison study of common NAD+ precursors in various organisms and mapped their biochemical roles in enhancing NAD+ levels. Moreover, we showed that increased NAD+ synthesis controls morphofunctional changes of activated macrophages, and directly imaged NAD+ declines during aging in situ. The broad utility of the FiNad sensors will expand our mechanistic understanding of numerous NAD+-associated physiological and pathological processes and facilitate screening for drug or gene candidates that affect uptake, efflux, and metabolism of this important cofactor.

Keywords: NAD(+) precursors; aging; fluorescence imaging; genetically encoded fluorescent sensor; nicotinamide adenine dinucleotide; real-time monitoring.

Conflict of interest statement

Declaration of Interests Y.Y., Y. Zhao, and Q.H. have filed a related patent by East China University of Science and Technology. The remaining authors declare no competing interests.

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