Engineering Fluorophore Recycling in a Fluorogenic RNA Aptamer. Academic Article uri icon

Overview

abstract

  • Fluorogenic aptamers can potentially show minimal photobleaching during continuous irradiation since any photobleached fluorophore can exchange with fluorescent dyes in the media. However, fluorophores have not been designed to maximize "fluorophore recycling." Here we describe TBI, a novel fluorophore for the Broccoli fluorogenic aptamer. Previous fluorophores either fail to rapidly dissociate when they undergo photobleaching via cis-trans isomerization, or bind slowly, resulting in extended periods after dissociation of the photobleached fluorophore when no fluorophore is bound. By contrast, photobleached TBI dissociates rapidly from Broccoli, and TBI from the media rapidly replaces dissociated photobleached fluorophore. Using TBI, Broccoli exhibits markedly enhanced fluorescence in cells during continuous imaging. These data show that designing fluorophores to optimize fluorophore recycling can lead to enhanced fluorescence of fluorogenic aptamers.

publication date

  • October 4, 2021

Research

keywords

  • Aptamers, Nucleotide
  • Fluorescent Dyes

Identity

PubMed Central ID

  • PMC8661118

Scopus Document Identifier

  • 85116464149

Digital Object Identifier (DOI)

  • 10.1002/anie.202108338

PubMed ID

  • 34490956

Additional Document Info

volume

  • 60

issue

  • 45