Efficient Reverse Transcription Using Locked Nucleic Acid Nucleotides towards the Evolution of Nuclease Resistant RNA Aptamers

M. Antopolsky; L. Crouzier; C. Dubois; S.L. Edwards; L.H. Lauridsen; J. Wengel; R.N. Veedu

doi:10.1371/journal.pone.0035990

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Efficient Reverse Transcription Using Locked Nucleic Acid Nucleotides towards the Evolution of Nuclease Resistant RNA Aptamers

Journal article

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Peer reviewed

Efficient Reverse Transcription Using Locked Nucleic Acid Nucleotides towards the Evolution of Nuclease Resistant RNA Aptamers

M. Antopolsky, L. Crouzier, C. Dubois, S.L. Edwards, L.H. Lauridsen, J. Wengel and R.N. Veedu

PloS one, Vol.7(4), e35990

2012

DOI: https://doi.org/10.1371/journal.pone.0035990

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Abstract

Background Modified nucleotides are increasingly being utilized in the de novo selection of aptamers for enhancing their drug-like character and abolishing the need for time consuming trial-and-error based post-selection modifications. Locked nucleic acid (LNA) is one of the most prominent and successful nucleic acid analogues because of its remarkable properties, and widely explored as building blocks in therapeutic oligonucleotides. Evolution of LNA-modified RNA aptamers requires an efficient reverse transcription method for PCR enrichment of the selected RNA aptamer candidates. Establishing this key step is a pre-requisite for performing LNA-modified RNA aptamer selection. Methodology In this study three different reverse transcriptases were investigated towards the enzymatic recognition of LNA nucleotides. Both incorporation as well as reading capabilities of the LNA nucleotides was investigated to fully understand the limitations of the enzymatic recognition. Conclusions We found that SuperScript® III Reverse Transcriptase is an efficient enzyme for the recognition of LNA nucleotides, making it a prime candidate to be used in de novo selection of LNA containing RNA aptamers

Details

Title: Efficient Reverse Transcription Using Locked Nucleic Acid Nucleotides towards the Evolution of Nuclease Resistant RNA Aptamers
Authors/Creators: M. Antopolsky (Author/Creator)
L. Crouzier (Author/Creator) - The University of Queensland
C. Dubois (Author/Creator) - The University of Queensland
S.L. Edwards (Author/Creator) - The University of Queensland
L.H. Lauridsen (Author/Creator) - Novo Nordisk Foundation
J. Wengel (Author/Creator) - University of Southern Denmark
R.N. Veedu (Author/Creator) - The University of Queensland
Publication Details: PloS one, Vol.7(4), e35990
Publisher: Public Library of Science
Identifiers: 991005543239507891
Copyright: © 2012 Crouzier et al.
Murdoch Affiliation: Murdoch University
Language: English
Resource Type: Journal article

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Collaboration types: Domestic collaboration; International collaboration
Citation topics: 2 Chemistry; 2.170 Nucleic Acids Chemistry; 2.170.988 Oligonucleotide Modifications
Web Of Science research areas: Biochemistry & Molecular Biology
ESI research areas: Chemistry