Dr. Jiesi Feng Profile

Dr. Jiesi Feng

at Peking Univ

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Proceedings Article | 1 March 2019 Presentation + Paper

G protein-assisted optimization of GPCR-activation based (GRAB) sensors

Zhaofa Wu, Jiesi Feng, Miao Jing, Yulong Li

Proceedings Volume 10865, 108650N (2019) https://doi.org/10.1117/12.2514631

KEYWORDS: Molecular biology, Fluorescent proteins, Proteins, Sensors, Life sciences, Brain, Neurotransmitters, In vivo imaging, Molecules

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In the brain, neurotransmitters or neuromodulators play pivotal roles in chemical synaptic transmission and consequently, monitoring their dynamics, especially in vivo, is critical for understanding their physiological- or pathophysiological roles at molecular, cellular, and circuit levels during behaviors and/or during diseases. We recently developed genetically-encoded GPCR-activation based (GRAB) sensors capable of reporting dynamics of acetylcholine, dopamine and norepinephrine with rapid kinetics, chemical- and cell-specificity in multiple organisms in vivo. Here, we explored the usage of G protein derivatives, either mini-G proteins or C-terminal peptides of Gα subunit to engineer new GRAB sensors. We found that the conformational changes mediated by mini-G proteins interacting with GPCRs, or Gα Cterminal peptides interacting with GPCRs could be harnessed to regulate fluorescence outputs of a GPCR fused circular permuted GFP (cpGFP). In addition, inter-molecular fusion of Gα C-terminal peptides significantly suppressed ectopic activation of G protein signaling in a GRAB acetylcholine sensor. Finally, we showed Gα C-terminal peptides fusion strategy could be applied to generate various GRAB sensors for small molecular compounds or neuropeptides.

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