Efficient Reverse Transcription Using Locked Nucleic Acid Nucleotides towards the Evolution of Nuclease Resistant RNA Aptamers
Identifieur interne : 005400 ( Main/Exploration ); précédent : 005399; suivant : 005401Efficient Reverse Transcription Using Locked Nucleic Acid Nucleotides towards the Evolution of Nuclease Resistant RNA Aptamers
Auteurs : Lucile Crouzier [Australie, France] ; Camille Dubois [Australie, France] ; Stacey L. Edwards [Australie] ; Lasse H. Lauridsen [Australie, Danemark] ; Jesper Wengel [Danemark] ; Rakesh N. Veedu [Australie]Source :
- PLoS ONE [ 1932-6203 ] ; 2012.
Abstract
Modified nucleotides are increasingly being utilized in the
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.
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
Url:
DOI: 10.1371/journal.pone.0035990
PubMed: 22558297
PubMed Central: 3338489
Affiliations:
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Edwards</name><affiliation wicri:level="1"><nlm:aff id="aff1"><addr-line>School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia, Brisbane, Queensland, Australia</addr-line></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea>School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia, Brisbane, Queensland</wicri:regionArea><wicri:noRegion>Queensland</wicri:noRegion></affiliation></author><author><name sortKey="Lauridsen, Lasse H" sort="Lauridsen, Lasse H" uniqKey="Lauridsen L" first="Lasse H." last="Lauridsen">Lasse H. Lauridsen</name><affiliation wicri:level="1"><nlm:aff id="aff1"><addr-line>School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia, Brisbane, Queensland, Australia</addr-line></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea>School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia, Brisbane, Queensland</wicri:regionArea><wicri:noRegion>Queensland</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff3"><addr-line>The Novo Nordisk Foundation Center for Biosustainability, Scion DTU, Hørsholm, Denmark</addr-line></nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>The Novo Nordisk Foundation Center for Biosustainability, Scion DTU, Hørsholm</wicri:regionArea><wicri:noRegion>Hørsholm</wicri:noRegion></affiliation></author><author><name sortKey="Wengel, Jesper" sort="Wengel, Jesper" uniqKey="Wengel J" first="Jesper" last="Wengel">Jesper Wengel</name><affiliation wicri:level="1"><nlm:aff id="aff4"><addr-line>Nucleic Acid Center, Department of Physics and Chemistry, University of Southern Denmark, Odense, Denmark</addr-line></nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Nucleic Acid Center, Department of Physics and Chemistry, University of Southern Denmark, Odense</wicri:regionArea><wicri:noRegion>Odense</wicri:noRegion></affiliation></author><author><name sortKey="Veedu, Rakesh N" sort="Veedu, Rakesh N" uniqKey="Veedu R" first="Rakesh N." last="Veedu">Rakesh N. Veedu</name><affiliation wicri:level="1"><nlm:aff id="aff1"><addr-line>School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia, Brisbane, Queensland, Australia</addr-line></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea>School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia, Brisbane, Queensland</wicri:regionArea><wicri:noRegion>Queensland</wicri:noRegion></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2012">2012</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background</title><p>Modified nucleotides are increasingly being utilized in the <italic>de novo</italic> 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.</p></sec><sec><title>Methodology</title><p>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.</p></sec><sec><title>Conclusions</title><p>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 <italic>de novo</italic> selection of LNA containing RNA aptamers</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Tuerk, C" uniqKey="Tuerk C">C Tuerk</name></author><author><name sortKey="Gold, L" uniqKey="Gold L">L Gold</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ellington, Ad" uniqKey="Ellington A">AD Ellington</name></author><author><name sortKey="Szostak, Jw" uniqKey="Szostak J">JW Szostak</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Famulok, M" uniqKey="Famulok M">M 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Lucile" uniqKey="Crouzier L" first="Lucile" last="Crouzier">Lucile Crouzier</name></noRegion><name sortKey="Dubois, Camille" sort="Dubois, Camille" uniqKey="Dubois C" first="Camille" last="Dubois">Camille Dubois</name><name sortKey="Edwards, Stacey L" sort="Edwards, Stacey L" uniqKey="Edwards S" first="Stacey L." last="Edwards">Stacey L. Edwards</name><name sortKey="Lauridsen, Lasse H" sort="Lauridsen, Lasse H" uniqKey="Lauridsen L" first="Lasse H." last="Lauridsen">Lasse H. Lauridsen</name><name sortKey="Veedu, Rakesh N" sort="Veedu, Rakesh N" uniqKey="Veedu R" first="Rakesh N." last="Veedu">Rakesh N. Veedu</name></country><country name="France"><region name="Hauts-de-France"><name sortKey="Crouzier, Lucile" sort="Crouzier, Lucile" uniqKey="Crouzier L" first="Lucile" last="Crouzier">Lucile Crouzier</name></region><name sortKey="Dubois, Camille" sort="Dubois, Camille" uniqKey="Dubois C" first="Camille" last="Dubois">Camille Dubois</name></country><country name="Danemark"><noRegion><name sortKey="Lauridsen, Lasse H" sort="Lauridsen, Lasse H" uniqKey="Lauridsen L" first="Lasse H." last="Lauridsen">Lasse H. Lauridsen</name></noRegion><name sortKey="Wengel, Jesper" sort="Wengel, Jesper" uniqKey="Wengel J" first="Jesper" last="Wengel">Jesper Wengel</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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