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Issue 12, 2017
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Evolved polymerases facilitate selection of fully 2′-OMe-modified aptamers

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Abstract

RNA or DNA aptamers with 2′-OMe-modifications have been pursued to increase resistance to nucleases, but have been difficult to identify because the OMe groups ablate polymerase recognition. We recently reported evolution of the thermostable DNA polymerases SFM4-6 and SFM4-9, which enable the efficient “transcription” and “reverse transcription”, respectively, of 2′-OMe oligonucleotides. With these polymerases, we now report the first selection of fully 2′-OMe modified aptamers, specifically aptamers that bind human neutrophil elastase (HNE). Two aptamers, 2mHNE-1 and 2mHNE-2, were isolated after five rounds of selection, and four more, 2mHNE-3–6, after an additional five rounds that included selection pressure for binding in the presence of serum. All six aptamers bind with reasonable affinity, which requires the 2′-OMe substituents. Further characterization of one aptamer, 2mHNE-5, showed that unlike a previously reported natural anti-HNE aptamer, affinity for HNE is retained in the presence of high concentrations of salt or serum. The polymerases SFM4-6 and SFM4-9 should prove valuable for the production and further exploration of modified aptamers.

Graphical abstract: Evolved polymerases facilitate selection of fully 2′-OMe-modified aptamers

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Supplementary files

Article information


Submitted
26 Aug 2017
Accepted
04 Oct 2017
First published
16 Oct 2017

This article is Open Access
All publication charges for this article have been paid for by the Royal Society of Chemistry

Chem. Sci., 2017,8, 8179-8182
Article type
Edge Article

Evolved polymerases facilitate selection of fully 2′-OMe-modified aptamers

Z. Liu, T. Chen and F. E. Romesberg, Chem. Sci., 2017, 8, 8179
DOI: 10.1039/C7SC03747C

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