Direct
Selection of Fluorescence-Enhancing RNA Aptamers
Version 2 2018-03-05, 16:18
Version 1 2018-02-13, 01:13
Posted on 2018-03-05 - 16:18
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 affinitya 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 ∼105 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. https://doi.org/10.1021/jacs.7b10724
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AUTHORS (6)
MG
Michael Gotrik
GS
Gurpreet Sekhon
SS
Saumya Saurabh
MN
Margaret Nakamoto
ME
Michael Eisenstein
HS
H. Tom Soh