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

Imai, S.; Hario, S.; Suerte, C.; Yamaguchi, I.; Sakoi, K.; Terai, T.; Takahashi-Yamashiro, K.; Campbell, R. E.

2025-04-17 molecular biology
10.1101/2025.04.17.649293 bioRxiv
Show abstract

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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