Optogenetics & Synthetic Biology Interdisciplinary Research Center, State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, Research Unit of Chinese Academy of Medical Sciences, East China University of Science and Technology, 130 Mei Long Road, Shanghai 200237, China; CAS Center for Excellence in Brain Science, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China; Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, 130 Mei Long Road, Shanghai 200237, China.
Optogenetics & Synthetic Biology Interdisciplinary Research Center, State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, Research Unit of Chinese Academy of Medical Sciences, East China University of Science and Technology, 130 Mei Long Road, Shanghai 200237, China; Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, 130 Mei Long Road, Shanghai 200237, China.
Dev Cell. 2020 Apr 20;53(2):240-252.e7. doi: 10.1016/j.devcel.2020.02.017. Epub 2020 Mar 19.
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.
对 NAD 代谢的理解为我们深入了解健康和疾病提供了许多关键的见解，但在活细胞和体内对 NAD 代谢进行高灵敏度和特异性的检测仍然很困难。在这里，我们提出了比率型、高响应的基因编码荧光指示剂 FiNad，用于监测活细胞和动物中的 NAD 动态。FiNad 传感器涵盖了生理相关的 NAD 浓度，并能灵敏地响应 NAD 的增加和减少。利用 FiNad，我们在各种生物体中对常见的 NAD 前体进行了头对头的比较研究，并绘制了它们在提高 NAD 水平方面的生化作用。此外，我们还表明，增加 NAD 的合成可以控制激活的巨噬细胞的形态和功能变化，并直接在体内原位成像衰老过程中 NAD 的下降。FiNad 传感器的广泛应用将扩展我们对许多与 NAD 相关的生理和病理过程的机制理解，并有助于筛选影响这种重要辅因子摄取、外排和代谢的药物或基因候选物。
