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Metabolite sensors

Fluorescent proteins (FPs) are widely used in many research elds, for example to track (sub)cellular localization of proteins and report gene expression activity. FPs can be also engineered to develop a sensor for real-time imaging of cellular events or activities. As they are genetically encoded proteins and self-suffcient to form intrinsic fluorophores without an extraneous chemical, it is possible to target these sensors to specific types of cells or even different subcellular organelles via signal peptides, thus allowing accurate spatiotemporal imaging of live-cell metabolic activities. To monitor intracellular events, researchers have developed genetically encoded biosensors for cellular metabolites, messengers, and conditions over the past two decades. These biosensors generally consist of two basic modules: substrate-binding proteins and FPs. From bacteria to mammals, various regulatory proteins and transcription factors specically sense intracellular biomolecules. The binding of biomolecules to the substrate-sensing protein often triggers conformational changes, which is transferred to the fused FP and affects the fluorescence intensity and/or spectra of the FP.

FR-biotechnology provides highly responsive, state of art genetically encoded sensors for monitoring cell metabolism in live cells or in vivo.


Frex, the Sensor specifically detect NADH

Frex series of genetically encoded fluorescent probes realize the dynamic detection and imaging of cell metabolism in various subcellular structures of living cells, and are powerful tools for studying cancer and metabolic diseases and drug screening. The picture shows the effect of mitochondrial inhibitors on NADH metabolism in intracellular mitochondria. (Cell Metabolism, 2011, 14, 555)

SoNar, a highly responsive sensor for measureing intracellular NADH/NAD+ ratio

SoNar is a highly responsive sensor for measureing intracellular or in vivo NADH/NAD+ ratio. SoNar enables dynamic monitoring and imaging of different types of cellular metabolic phenotypes in live cells and live animals, as well as high-throughput screening of active compounds associated with cell metabolism. We found specific metabolic patterns of cancer stem cells, and also found some significant changes in cell metabolism of anti-cancer drugs(Cell Metabolism 2015, 21, 777; Nature Protocols, 2016, 11, 1345; Cell Metabolism 2019, 29, 950;Blood 2020, 136, 553)

iNap, highly responsive sensors reveal intracellular NADPH dynamics

iNap sensors are highly specific iNADPH sensors with various affinity. iNaps enable high-spatiotemporal resolution detection and imaging of NADPH metabolism in vivo, live cells and various subcellular structures, and can be used for the study of antioxidant, biosynthesis, AMPK and other pathways (Nature Methods, 2017, 14, 720; Nature Protocols, 2018)

FiNad, high responsive sensor for NAD+ Metabolism in Live Cells and In Vivo 

FiNad is a highly responsive, sensitive, and large dynamic range genetically encoded fluorescent probe for detecting NAD+/AXP ratios, enabling small-level NAD+ imaging of bacteria, yeast, mammalian cells, zebrafish, and living organisms. , can be used for cell signal transduction, cell metabolism, aging and other fields research (Developmental Cell, 2020)

FiLa: ultrasensitive lactate sensors reveal the spatiotemporal landscape of lactate metabolism in physiology and disease

Lactate is an important energy fuel, synthetic building block and signaling molecule, and is a metabolic "star" with multiple key roles, playing an important role in physiological and pathological processes. Lactic acid metabolism presents drastic dynamic changes and complex spatial distribution, and traditional biochemical methods are difficult to achieve dynamic tracking at the living cell and in vivo levels. FiLa is a highly specific, highly responsive, ultrasensitive lactic acid fluorescent probe, to map subcellular lactate metabolism, and this study accidentally found high concentrations of lactic acid enriched in the mitochondrial matrix, thus concluded important scientific problems of mitochondrial lactate metabolism that have been debated in this field for decades. A point-of-care detection technology for clinical body fluid samples based on FiLa probe was established, and a significant increase in urine lactate was found to be a potential screening marker for maternal hereditary diabetes mellitus with deafness (MIDD) disease, which provided an important basis for clinical diagnosis. (Cell Metabolism 2023, 35, 220)



  1. Li, X. et al. Ultrasensitive sensors reveal the spatiotemporal landscape of lactate metabolism in physiology and disease. Cell metabolism 35, 200-211 e209, doi:10.1016/j.cmet.2022.10.002 (2023).
  2. Chen, C. et al. Oxidative phosphorylation enhances the leukemogenic capacity and resistance to chemotherapy of B cell acute lymphoblastic leukemia. Sci Adv 7, eabd6280, doi:10.1126/sciadv.abd6280 (2021).
  3. Zou, Y. et al. Illuminating NAD+ Metabolism in Live Cells and In Vivo Using a Genetically Encoded Fluorescent Sensor. Developmental cell 53, 240-252, doi: (2020).
  4. Zhang, Z., Cheng, X., Zhao, Y. & Yang, Y. Lighting Up Live-Cell and In Vivo Central Carbon Metabolism with Genetically Encoded Fluorescent Sensors. Annu Rev Anal Chem 13, 293-314, doi:10.1146/annurev-anchem-091619-091306 (2020).
  5. Lim, S. L. et al. In planta study of photosynthesis and photorespiration using NADPH and NADH/NAD(+) fluorescent protein sensors. Nature communications 11, 3238, doi:10.1038/s41467-020-17056-0 (2020).
  6. Gu, H. et al. MDH1-mediated malate-aspartate NADH shuttle maintains the activity levels of fetal liver hematopoietic stem cells. Blood 136, 553-571, doi:10.1182/blood.2019003940 (2020).
  7. Hao, X. et al. Metabolic Imaging Reveals a Unique Preference of Symmetric Cell Division and Homing of Leukemia-Initiating Cells in an Endosteal Niche. Cell metabolism 29, 950-965, doi:10.1016/j.cmet.2018.11.013 (2019).
  8. Zou, Y. et al. Analysis of redox landscapes and dynamics in living cells and in vivo using genetically encoded fluorescent sensors. Nat Protoc 13, 2362-2386, doi:10.1038/s41596-018-0042-5 (2018).
  9. Zhao, Y., Zhang, Z., Zou, Y. & Yang, Y. Visualization of nicotine adenine dinucleotides redox homeostasis with genetically encoded fluorescent sensors. Redox. Signal. 28, 213-229, doi:10.1089/ars.2017.7226 (2018).
  10. Liu, X. et al. PPM1K Regulates Hematopoiesis and Leukemogenesis through CDC20-Mediated Ubiquitination of MEIS1 and p21. Cell reports 23, 1461-1475, doi:10.1016/j.celrep.2018.03.140 (2018).
  11. Tao, R. et al. Genetically encoded fluorescent sensors reveal dynamic regulation of NADPH metabolism. Nature methods 14, 720-728, doi:10.1038/nmeth.4306 (2017).
  12. Zhao, Y. et al. In vivo monitoring of cellular energy metabolism using a highly responsive sensor for NAD+/NADH redox state. Nature Protocols 11, 1345-1359 (2016).
  13. Zhao, Y. et al. SoNar, a Highly Responsive NAD(+)/NADH Sensor, Allows High-Throughput Metabolic Screening of Anti-tumor Agents. Cell metabolism 21, 777-789, doi:10.1016/j.cmet.2015.04.009 (2015).
  14. Zhao, Y. et al. Genetically Encoded Fluorescent Sensors for Intracellular NADH Detection. Cell metabolism 14, 555-566, doi:10.1016/j.cmet.2011.09.004 (2011).