RNA Mango Aptamer-Fluorophore: A Bright, High-Affinity
Complex for RNA Labeling and Tracking
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Abstract
Because RNA lacks strong intrinsic
fluorescence, it has proven
challenging to track RNA molecules in real time. To address this problem
and to allow the purification of fluorescently tagged RNA complexes,
we have selected a high affinity RNA aptamer called RNA Mango. This
aptamer binds a series of thiazole orange (fluorophore) derivatives
with nanomolar affinity, while increasing fluorophore fluorescence
by up to 1,100-fold. Visualization of RNA Mango by single-molecule
fluorescence microscopy, together with injection and imaging of RNA
Mango/fluorophore complex in <i>C. elegans</i> gonads demonstrates
the potential for live-cell RNA imaging with this system. By inserting
RNA Mango into a stem loop of the bacterial 6S RNA and biotinylating
the fluorophore, we demonstrate that the aptamer can be used to simultaneously
fluorescently label and purify biologically important RNAs. The high
affinity and fluorescent properties of RNA Mango are therefore expected
to simplify the study of RNA complexes