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April 21st, 2025
Version: 2
The University of Tokyo
molecular biology
biorxiv

High-performance genetically-encoded green and red fluorescent biosensors for pyruvate

Imai, S.Open in Google Scholar•Hario, S.Open in Google Scholar•Suerte, C.Open in Google Scholar•Yamaguchi, I.Open in Google Scholar•Sakoi, K.Open in Google Scholar•Terai, T.Open in Google Scholar•Takahashi-Yamashiro, K.Open in Google Scholar•Campbell, R. E.Open in Google Scholar

Pyruvate is the end-product of glycolysis and a central metabolite involved in many biochemical pathways. However, a lack of high-performance (i.e., {Delta}F/F0 > 10) single fluorescent protein (FP)-based biosensors has hindered efforts to investigate the physiological role of pyruvate. Here, we present the GreenPy1 and ApplePy1 series, which are green FP (GFP)-based and red FP (RFP)-based pyruvate biosensors, respectively. Both series exhibit large fluorescence intensity change ({Delta}F/F0 [~] 20 to >40) and a range of affinities (10s of M to several mM). We demonstrate the utility of these pyruvate biosensors for multicolor imaging of metabolite concentration changes in mammalian cells.

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