Efficient reverse transcription using locked Nucleic Acid Nucleotides towards the evolution of Nuclease resistant RNA Aptamers

Crouzier, Lucile, Dubois, Camille, Edwards, Stacey L., Lauridsen, Lasse H., Wengel, Jesper and Veedu, Rakesh N. (2012) Efficient reverse transcription using locked Nucleic Acid Nucleotides towards the evolution of Nuclease resistant RNA Aptamers. PLoS One, 7 4: e35990-1-e35990-5. doi:10.1371/journal.pone.0035990


Author Crouzier, Lucile
Dubois, Camille
Edwards, Stacey L.
Lauridsen, Lasse H.
Wengel, Jesper
Veedu, Rakesh N.
Title Efficient reverse transcription using locked Nucleic Acid Nucleotides towards the evolution of Nuclease resistant RNA Aptamers
Journal name PLoS One   Check publisher's open access policy
ISSN 1932-6203
Publication date 2012-04-25
Sub-type Article (original research)
DOI 10.1371/journal.pone.0035990
Open Access Status DOI
Volume 7
Issue 4
Start page e35990-1
End page e35990-5
Total pages 5
Place of publication San Francisco, CA, United States
Publisher Public Library of Science
Collection year 2013
Language eng
Formatted 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
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2013 Collection
School of Chemistry and Molecular Biosciences
 
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Created: Tue, 08 May 2012, 19:04:32 EST by Lucy O'Brien on behalf of School of Chemistry & Molecular Biosciences