Fluorophore Binding Aptamers as a Tool for RNA Visualization


By jpeza - Posted on 29 Junio 2009

Fecha Publicación: 
1 May 2009
Nombre de Revista: 
Datos del paper
Autor Principal: 
Katja Eydeler
Volumen: 
96
Issue: 
6
Página Inicial: 
3703
Página Final: 
3707
Abstract: 

Fluorescence correlation spectroscopy (FCS) is suitable for the detection of fluorescent molecules in living cells.

For the visualization of mRNA, we genetically fused a fluorophore-specific RNA aptamer to the coding mRNA of the green fluorescent

protein, as well as to noncoding sequences. Using these constructs, we showed that the aptamer portion of the mRNA

still binds the fluorophore in the nanomolar range as determined via FCS. Furthermore, the binding took place in the context of

total RNA extract. A tandem construct of the RNA aptamer even exhibited a lower Kd than the monomer. This FCS-based method

establishes a tool for minimal invasive detection of RNA at the single molecule level in individual living cells.

Dirección del Autor: 

Department of Chemistry, Institute of Biochemistry and Molecular Biology, University of Hamburg, 20146 Hamburg, Germany

Keywords: 
Fluorescence correlation spectroscopy ; coding mRNA ; mRNA
Coautores: 

Eileen Magbanua; Arne Werner, Patrick Ziegelmuller ;  Ulrich Hahn

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