Lactate biosensors for spectrally and spatially multiplexed fluorescence imaging

Yusuke Nasu (), Abhi Aggarwal, Giang N. T. Le, Camilla Trang Vo, Yuki Kambe, Xinxing Wang, Felix R. M. Beinlich, Ashley Bomin Lee, Tina R. Ram, Fangying Wang, Kelsea A. Gorzo, Yuki Kamijo, Marc Boisvert, Suguru Nishinami, Genki Kawamura, Takeaki Ozawa, Hirofumi Toda, Grant R. Gordon, Shaoyu Ge, Hajime Hirase, Maiken Nedergaard, Marie-Eve Paquet, Mikhail Drobizhev, Kaspar Podgorski and Robert E. Campbell ()
Additional contact information
Yusuke Nasu: The University of Tokyo, Bunkyo-ku
Abhi Aggarwal: Janelia Research Campus, Howard Hughes Medical Institute
Giang N. T. Le: The University of Tokyo, Bunkyo-ku
Camilla Trang Vo: University of Copenhagen
Yuki Kambe: Kagoshima University, Sakuragaoka
Xinxing Wang: Stony Brook University
Felix R. M. Beinlich: University of Copenhagen
Ashley Bomin Lee: University of Copenhagen
Tina R. Ram: University of Calgary
Fangying Wang: University of Calgary
Kelsea A. Gorzo: University of Calgary
Yuki Kamijo: The University of Tokyo, Bunkyo-ku
Marc Boisvert: CERVO Brain Research Centre
Suguru Nishinami: University of Tsukuba, Tsukuba
Genki Kawamura: The University of Tokyo, Bunkyo-ku
Takeaki Ozawa: The University of Tokyo, Bunkyo-ku
Hirofumi Toda: University of Tsukuba, Tsukuba
Grant R. Gordon: University of Calgary
Shaoyu Ge: Stony Brook University
Hajime Hirase: University of Copenhagen
Maiken Nedergaard: University of Copenhagen
Marie-Eve Paquet: CERVO Brain Research Centre
Mikhail Drobizhev: Montana State University
Kaspar Podgorski: Janelia Research Campus, Howard Hughes Medical Institute
Robert E. Campbell: The University of Tokyo, Bunkyo-ku

Nature Communications, 2023, vol. 14, issue 1, 1-17

Abstract: Abstract l-Lactate is increasingly appreciated as a key metabolite and signaling molecule in mammals. However, investigations of the inter- and intra-cellular dynamics of l-lactate are currently hampered by the limited selection and performance of l-lactate-specific genetically encoded biosensors. Here we now report a spectrally and functionally orthogonal pair of high-performance genetically encoded biosensors: a green fluorescent extracellular l-lactate biosensor, designated eLACCO2.1, and a red fluorescent intracellular l-lactate biosensor, designated R-iLACCO1. eLACCO2.1 exhibits excellent membrane localization and robust fluorescence response. To the best of our knowledge, R-iLACCO1 and its affinity variants exhibit larger fluorescence responses than any previously reported intracellular l-lactate biosensor. We demonstrate spectrally and spatially multiplexed imaging of l-lactate dynamics by coexpression of eLACCO2.1 and R-iLACCO1 in cultured cells, and in vivo imaging of extracellular and intracellular l-lactate dynamics in mice.

Date: 2023
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DOI: 10.1038/s41467-023-42230-5

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