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A Fluorogenic RNA-Based Sensor Activated by Metabolite-Induced RNA Dimerization.

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Cell Chemical Biology
|October 22, 2019
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Summary
This summary is machine-generated.

Researchers developed a novel RNA biosensor to detect S-adenosylmethionine (SAM) by regulating the dimerization of the Corn aptamer. This advancement enables real-time monitoring of SAM in living cells, overcoming photobleaching limitations.

Keywords:
RNA dimerizationS-adenosylmethionine (SAM) riboswitchcell imagingfluorogenic aptamermetabolite dynamics

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • RNA Aptamer Technology

Background:

  • The Corn aptamer exhibits high-affinity homodimerization and binds DFHO, producing photostable fluorescence.
  • Photobleaching limits the quantitative accuracy of RNA-based biosensors.
  • Developing regulated biosensors is crucial for monitoring small molecules in biological systems.

Purpose of the Study:

  • To engineer a small-molecule-regulated fluorescent biosensor using the Corn aptamer.
  • To detect S-adenosylmethionine (SAM) in vitro and in living cells.
  • To create a novel class of RNA-based biosensors based on regulated aptamer dimerization.

Main Methods:

  • Fused the Corn aptamer with a circularly permuted SAM aptamer from the SAM-III riboswitch.
  • Engineered a helical stem to control Corn dimerization.
  • Assessed biosensor function in vitro and in living mammalian cells.

Main Results:

  • The engineered fusion RNA functions as a SAM-dependent biosensor.
  • In the absence of SAM, Corn fails to dimerize; upon SAM binding, dimerization and DFHO binding occur.
  • Successfully detected SAM dynamics in living mammalian cells.

Conclusions:

  • A novel strategy for converting constitutive aptamers into regulated biosensors was established.
  • The developed RNA biosensor enables sensitive detection of SAM dynamics in live cells.
  • This work introduces a new platform for RNA-based biosensors utilizing small-molecule-regulated aptamer dimerization.