Turning an Aptamer into a Light-Switch Probe with a Single Bioconjugation
Identifieur interne : 000225 ( Main/Exploration ); précédent : 000224; suivant : 000226Turning an Aptamer into a Light-Switch Probe with a Single Bioconjugation
Auteurs : Thakshila X0a M. Wickramaratne ; Valerie C. PierreSource :
- Bioconjugate Chemistry [ 1043-1802 ] ; 2014.
Abstract
We describe a method for transforming a structure-switching aptamer into a luminescent light-switch probe via a single conjugation. The methodology is demonstrated using a known aptamer for Hg2+ as a case study. This approach utilizes a lanthanide-based metallointercalator, Eu-DOTA-Phen, whose luminescence is quenched almost entirely and selectively by purines, but not at all by pyrimidines. This complex, therefore, does not luminesce while intercalated in dsDNA, but it is bright red when conjugated to a ssDNA that is terminated by several pyrimidines. In its design, the light-switch probe incorporates a structure-switching aptamer partially hybridized to its complementary strand. The lanthanide complex is conjugated to either strand via a stable amide bond. Binding of the analyte by the structure-switching aptamer releases the complementary strand. This release precludes intercalation of the intercalator in dsDNA, which switches on its luminescence. The resulting probe turns on 21-fold upon binding to its analyte. Moreover, the structure switching aptamer is highly selective, and the long luminescence lifetime of the probe readily enables time-gating experiments for removal of the background autofluorescence of the sample.
Url:
DOI: 10.1021/bc5003899
PubMed: 25427946
PubMed Central: 4306522
Affiliations:
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Turning an Aptamer into a Light-Switch Probe with
a Single Bioconjugation</title><author><name sortKey="Wickramaratne, Thakshila X0a M" sort="Wickramaratne, Thakshila X0a M" uniqKey="Wickramaratne T" first="Thakshila X0a M." last="Wickramaratne">Thakshila X0a M. Wickramaratne</name></author><author><name sortKey="Pierre, Valerie C" sort="Pierre, Valerie C" uniqKey="Pierre V" first="Valerie C." last="Pierre">Valerie C. Pierre</name></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">25427946</idno><idno type="pmc">4306522</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4306522</idno><idno type="RBID">PMC:4306522</idno><idno type="doi">10.1021/bc5003899</idno><date when="2014">2014</date><idno type="wicri:Area/Pmc/Corpus">000071</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000071</idno><idno type="wicri:Area/Pmc/Curation">000071</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000071</idno><idno type="wicri:Area/Pmc/Checkpoint">000097</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000097</idno><idno type="wicri:Area/Ncbi/Merge">000925</idno><idno type="wicri:Area/Ncbi/Curation">000925</idno><idno type="wicri:Area/Ncbi/Checkpoint">000925</idno><idno type="wicri:doubleKey">1043-1802:2014:Wickramaratne T:turning:an:aptamer</idno><idno type="wicri:Area/Main/Merge">000225</idno><idno type="wicri:Area/Main/Curation">000225</idno><idno type="wicri:Area/Main/Exploration">000225</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Turning an Aptamer into a Light-Switch Probe with
a Single Bioconjugation</title><author><name sortKey="Wickramaratne, Thakshila X0a M" sort="Wickramaratne, Thakshila X0a M" uniqKey="Wickramaratne T" first="Thakshila X0a M." last="Wickramaratne">Thakshila X0a M. Wickramaratne</name></author><author><name sortKey="Pierre, Valerie C" sort="Pierre, Valerie C" uniqKey="Pierre V" first="Valerie C." last="Pierre">Valerie C. Pierre</name></author></analytic><series><title level="j">Bioconjugate Chemistry</title><idno type="ISSN">1043-1802</idno><idno type="eISSN">1520-4812</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p content-type="toc-graphic"><graphic xlink:href="bc-2014-003899_0009" id="ab-tgr1"></graphic></p><p>We
describe a method for transforming a structure-switching aptamer
into a luminescent light-switch probe via a single conjugation. The
methodology is demonstrated using a known aptamer for Hg<sup>2+</sup> as a case study. This approach utilizes a lanthanide-based metallointercalator,
Eu-DOTA-Phen, whose luminescence is quenched almost entirely and selectively
by purines, but not at all by pyrimidines. This complex, therefore,
does not luminesce while intercalated in dsDNA, but it is bright red
when conjugated to a ssDNA that is terminated by several pyrimidines.
In its design, the light-switch probe incorporates a structure-switching
aptamer partially hybridized to its complementary strand. The lanthanide
complex is conjugated to either strand via a stable amide bond. Binding
of the analyte by the structure-switching aptamer releases the complementary
strand. This release precludes intercalation of the intercalator in
dsDNA, which switches on its luminescence. The resulting probe turns
on 21-fold upon binding to its analyte. Moreover, the structure switching
aptamer is highly selective, and the long luminescence lifetime of
the probe readily enables time-gating experiments for removal of the
background autofluorescence of the sample.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Liu, J W" uniqKey="Liu J">J. W. Liu</name></author><author><name sortKey="Cao, Z H" uniqKey="Cao Z">Z. H. Cao</name></author><author><name sortKey="Lu, Y" uniqKey="Lu Y">Y. Lu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Stojanovic, M N" uniqKey="Stojanovic M">M. N. Stojanovic</name></author><author><name sortKey="De Prada, P" uniqKey="De Prada P">P. de Prada</name></author><author><name sortKey="Landry, D W" uniqKey="Landry D">D. W. Landry</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Stojanovic, M N" uniqKey="Stojanovic M">M. N. Stojanovic</name></author><author><name sortKey="De Prada, P" uniqKey="De Prada P">P. de Prada</name></author><author><name sortKey="Landry, D W" uniqKey="Landry D">D. W. 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