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RIBOsensor for FRET-based, real-time ribose measurements in live cells.

Mina Ahmadi1, Zhuangyu Zhao1, Ivan J Dmochowski1

  • 1Department of Chemistry, University of Pennsylvania Philadelphia PA USA ivandmo@sas.upenn.edu.

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Scientists developed a new sensor to track d-ribose, a key molecule for energy and nucleic acids. This tool helps study ribose dynamics in cells, crucial for understanding energy metabolism in health and disease.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Background:

  • d-Ribose is a fundamental building block of nucleic acids and ATP.
  • Cellular ribose distribution, biosynthesis, and regulation are not well understood.
  • Existing methods for ribose detection are limited for live-cell imaging.

Purpose of the Study:

  • To develop a novel biosensor for real-time monitoring of intracellular d-ribose dynamics.
  • To enable quantitative analysis of ribose transport and metabolism in live mammalian cells.
  • To provide a tool for investigating ribose's role in cellular energy production under various conditions.

Main Methods:

  • Genetically encoded fluorescence resonance energy transfer (FRET) sensor construction using ribose-binding protein (RBP) and fluorescent proteins (FPs).
  • Optimized FP placement near the RBP active site to enhance FRET signal upon ribose binding.
  • Validation of the sensor's sensitivity, reversibility, and selectivity in live-cell imaging experiments.

Main Results:

  • The developed RIBOsensor exhibits increased FRET signal upon d-ribose binding.
  • The sensor allows for rapid, reversible, and selective detection of labile ribose in live cells.
  • The sensor enables longitudinal studies and intracellular ribose quantitation.

Conclusions:

  • The RIBOsensor is a sensitive and specific tool for monitoring intracellular ribose.
  • This technology facilitates the investigation of ribose metabolism and transport in physiological and pathological states.
  • Understanding ribose dynamics can offer insights into cellular energy production and disease mechanisms.