@article{6de2d6c506ae4edf9c7779ce48f3abcb,
title = "A high-performance genetically encoded fluorescent indicator for in vivo cAMP imaging",
abstract = "cAMP is a key second messenger that regulates diverse cellular functions including neural plasticity. However, the spatiotemporal dynamics of intracellular cAMP in intact organisms are largely unknown due to low sensitivity and/or brightness of current genetically encoded fluorescent cAMP indicators. Here, we report the development of the new circularly permuted GFP (cpGFP)-based cAMP indicator G-Flamp1, which exhibits a large fluorescence increase (a maximum ΔF/F0 of 1100% in HEK293T cells), decent brightness, appropriate affinity (a Kd of 2.17 μM) and fast response kinetics (an association and dissociation half-time of 0.20 and 0.087 s, respectively). Furthermore, the crystal structure of the cAMP-bound G-Flamp1 reveals one linker connecting the cAMP-binding domain to cpGFP adopts a distorted β-strand conformation that may serve as a fluorescence modulation switch. We demonstrate that G-Flamp1 enables sensitive monitoring of endogenous cAMP signals in brain regions that are implicated in learning and motor control in living organisms such as fruit flies and mice.",
author = "Liang Wang and Chunling Wu and Wanling Peng and Ziliang Zhou and Jianzhi Zeng and Xuelin Li and Yini Yang and Shuguang Yu and Ye Zou and Mian Huang and Chang Liu and Yefei Chen and Yi Li and Panpan Ti and Wenfeng Liu and Yufeng Gao and Wei Zheng and Haining Zhong and Shangbang Gao and Zhonghua Lu and Ren, {Pei Gen} and Ng, {Ho Leung} and Jie He and Shoudeng Chen and Min Xu and Yulong Li and Jun Chu",
note = "Funding Information: This work was supported by National Key Research and Development Program of China (2021YFF0502904 to J.C., 2020YFA0908802 to L.W., 2017YFA0700403 to J.C.), National Natural Science Foundation of China (81927803 and 21874145 to J.C., 32000732 to L.W.), Guangdong Basic and Applied Basic Research Foundation (2020B121201010 to J.C.), Natural Science Foundation of Shenzhen (JCYJ20200109115633343 to J.C.), CAS grants (DWKF20200001 and NSY889021058 to J.C.). We thank Drs. Michael Lin at Stanford University, Jonathan A. Cooper at Fred Hutchinson Cancer Research Center, Fran{\c c}ois St-Pierre at Baylor College of Medicine, Yu Mu at Institute of Neuroscience-CAS, Yulin Zhao at Peking University, and Fang Liu at the Campbell Family Mental Health Research Institute, for the critical reading of this manuscript. Funding Information: This work was supported by National Key Research and Development Program of China (2021YFF0502904 to J.C., 2020YFA0908802 to L.W., 2017YFA0700403 to J.C.), National Natural Science Foundation of China (81927803 and 21874145 to J.C., 32000732 to L.W.), Guangdong Basic and Applied Basic Research Foundation (2020B121201010 to J.C.), Natural Science Foundation of Shenzhen (JCYJ20200109115633343 to J.C.), CAS grants (DWKF20200001 and NSY889021058 to J.C.). We thank Drs. Michael Lin at Stanford University, Jonathan A. Cooper at Fred Hutchinson Cancer Research Center, Fran{\c c}ois St-Pierre at Baylor College of Medicine, Yu Mu at Institute of Neuroscience-CAS, Yulin Zhao at Peking University, and Fang Liu at the Campbell Family Mental Health Research Institute, for the critical reading of this manuscript. Publisher Copyright: {\textcopyright} 2022, The Author(s).",
year = "2022",
month = dec,
doi = "10.1038/s41467-022-32994-7",
language = "English (US)",
volume = "13",
journal = "Nature Communications",
issn = "2041-1723",
publisher = "Nature Publishing Group",
number = "1",
}