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Determination of In Vitro and Cellular Turn-on Kinetics for Fluorogenic RNA Aptamers
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Engineering Fluorophore Recycling in a Fluorogenic RNA Aptamer.

Xing Li1,2, Jiahui Wu2, Samie R Jaffrey2

  • 1Beijing Institutes of Life Science, Chinese Academy of Sciences, Beijing, 100101, P. R. China.

Angewandte Chemie (International Ed. in English)
|September 7, 2021
PubMed
Summary
This summary is machine-generated.

A novel fluorophore, TBI, enhances fluorescence in Broccoli aptamers by enabling rapid "fluorophore recycling." This design overcomes limitations of previous fluorophores, improving continuous imaging in cells.

Keywords:
RNA imagingfluorescent probesfluorophore recyclingphotostability

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Chemical Biology

Background:

  • Fluorogenic aptamers offer potential for minimal photobleaching due to fluorophore exchange.
  • Current fluorophores are not optimized for efficient "fluorophore recycling."

Purpose of the Study:

  • To introduce TBI, a novel fluorophore designed to maximize fluorophore recycling for the Broccoli aptamer.
  • To address limitations of existing fluorophores, such as slow binding and dissociation after photobleaching.

Main Methods:

  • Design and synthesis of the novel fluorophore TBI.
  • Characterization of TBI's photobleaching and dissociation kinetics with the Broccoli aptamer.
  • In-cell imaging experiments to assess fluorescence enhancement during continuous irradiation.

Main Results:

  • TBI demonstrates rapid dissociation from Broccoli after photobleaching.
  • TBI from the media efficiently replaces photobleached fluorophores, minimizing "dark" periods.
  • Broccoli aptamer exhibits significantly enhanced fluorescence in cells when using TBI during continuous imaging.

Conclusions:

  • Designing fluorophores for optimized fluorophore recycling enhances the performance of fluorogenic aptamers.
  • TBI represents a significant advancement for continuous imaging applications using aptamer-based fluorescent sensors.