Direct Selection of Fluorescence-Enhancing RNA Aptamers

Published on 2018-03-05T16:18:33Z (GMT) by
RNA aptamers that generate a strong fluorescence signal upon binding a nonfluorescent small-molecule dye offer a powerful means for the selective imaging of individual RNA species. Unfortunately, conventional in vitro discovery methods are not efficient at generating such fluorescence-enhancing aptamers, because they primarily exert selective pressure based on target affinitya characteristic that correlates poorly with fluorescence enhancement. Thus, only a handful of fluorescence-enhancing aptamers have been reported to date. In this work, we describe a method for converting DNA libraries into “gene-linked RNA aptamer particles” (GRAPs) that each display ∼10<sup>5</sup> copies of a single RNA sequence alongside the DNA that encodes it. We then screen large libraries of GRAPs in a high-throughput manner using the FACS instrument based directly on their fluorescence-enhancing properties. Using this strategy, we demonstrate the capability to generate fluorescence-enhancing aptamers that produce a variety of different emission wavelengths upon binding the dye of interest.

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Gotrik, Michael; Sekhon, Gurpreet; Saurabh, Saumya; Nakamoto, Margaret; Eisenstein, Michael; Soh, H. Tom (2018): Direct

Selection of Fluorescence-Enhancing RNA Aptamers. ACS Publications. Collection.