bioRxivBioengineeringPreprint
High performance genetically-encoded green fluorescent biosensors for intracellular L-lactate
G. N. T. Le, S. Hario, K. Takahashi-Yamashiro, S. Li, M. Drobizhev, Y. Nasu, R. E. Campbell
Abstract
L-Lactate is a monocarboxylate produced during the process of cellular glycolysis and has long been generally considered a waste product. However, studies in recent decades have provided new perspectives on the physiological roles of L-lactate as a major energy substrate and a signaling molecule. To enable further investigations of the physiological roles of L-lactate, we have developed a series of high-performance ({Delta}F/F = 15 to 30 in vitro), intensiometric, genetically-encoded green fluorescent protein (GFP)-based intracellular L-lactate biosensors with a range of affinities. We evaluated the performance of these biosensors by in vitro and live-cell characterization and demonstrated the utility with imaging applications in several cell lines.
- assay: fluorescence / live-cell microscopy
- research focus: protein structure prediction
- approach: experimental assay
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