A high-performance genetically encoded fluorescent indicator for in vivo cAMP imaging

Liang Wang, Chunling Wu, Wanling Peng, Ziliang Zhou, Jianzhi Zeng, Xuelin Li, Yini Yang, Shuguang Yu, Ye Zou, Mian Huang, Chang Liu, Yefei Chen, Yi Li, Panpan Ti, Wenfeng Liu, Yufeng Gao, Wei Zheng, Haining Zhong, Shangbang Gao, Zhonghua Lu, Pei-Gen Ren, Ho Leung Ng, Jie He, Shoudeng Chen, Min Xu, Yulong Li and Jun Chu ()
Additional contact information
Liang Wang: Chinese Academy of Sciences
Chunling Wu: PKU-IDG–McGovern Institute for Brain Research
Wanling Peng: Chinese Academy of Sciences
Ziliang Zhou: Sun Yat-sen University
Jianzhi Zeng: PKU-IDG–McGovern Institute for Brain Research
Xuelin Li: PKU-IDG–McGovern Institute for Brain Research
Yini Yang: PKU-IDG–McGovern Institute for Brain Research
Shuguang Yu: Chinese Academy of Sciences
Ye Zou: Kansas State University
Mian Huang: Kansas State University
Chang Liu: Chinese Academy of Sciences
Yefei Chen: Chinese Academy of Sciences
Yi Li: Huazhong University of Science and Technology
Panpan Ti: Huazhong University of Science and Technology
Wenfeng Liu: Chinese Academy of Sciences
Yufeng Gao: Chinese Academy of Sciences
Wei Zheng: Chinese Academy of Sciences
Haining Zhong: Oregon Health and Science University
Shangbang Gao: Huazhong University of Science and Technology
Zhonghua Lu: Chinese Academy of Sciences
Pei-Gen Ren: Chinese Academy of Sciences
Ho Leung Ng: Kansas State University
Jie He: Chinese Academy of Sciences
Shoudeng Chen: Sun Yat-sen University
Min Xu: Chinese Academy of Sciences
Yulong Li: PKU-IDG–McGovern Institute for Brain Research
Jun Chu: Chinese Academy of Sciences

Nature Communications, 2022, vol. 13, issue 1, 1-16

Abstract: 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.

Date: 2022
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DOI: 10.1038/s41467-022-32994-7

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