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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
10
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pubmed:dateCreated |
2009-9-15
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pubmed:abstractText |
An RNA editing reaction that is both essential and specific to the trypanosomatid parasites is an attractive target for new drug development. Although high-throughput screening of chemical libraries is a powerful strategy often used to identify new drugs, the available in vitro editing assays do not have the necessary sensitivity and format for this approach to be feasible. A ruthenium labeled reporter RNA is described here that overcomes these limitations as it can both detect edited product in the low femtomole range and is ideal for high-throughput format. The reporter RNA consists of an RNA editing substrate linked to a streptavidin-binding aptamer that is initially held within an inactive conformation. An in vitro selection strategy optimized the linkage so that the streptavidin-binding aptamer is only activated by an editing-induced conformational change. An electrochemiluminescent signal results from the ruthenium label when the reporter is bound to the bottom of a streptavidin-coated microtiter plate where it can be stimulated by a carbon electrode. Chemical probing, mutagenesis, and binding affinity measurements were used to characterize the reporter. The highly sensitive assay could be adapted to a broad range of RNA processing reactions.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/19696159-10037718,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19696159-10074372,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/19696159-9799244
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Oct
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pubmed:issn |
1469-9001
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pubmed:author |
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pubmed:issnType |
Electronic
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pubmed:volume |
15
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1929-38
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pubmed:dateRevised |
2010-9-27
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pubmed:meshHeading |
pubmed-meshheading:19696159-Aptamers, Nucleotide,
pubmed-meshheading:19696159-Base Sequence,
pubmed-meshheading:19696159-Electrochemistry,
pubmed-meshheading:19696159-Luminescence,
pubmed-meshheading:19696159-Molecular Sequence Data,
pubmed-meshheading:19696159-Mutation,
pubmed-meshheading:19696159-Nucleic Acid Conformation,
pubmed-meshheading:19696159-RNA Editing,
pubmed-meshheading:19696159-Sequence Homology, Nucleic Acid
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pubmed:year |
2009
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pubmed:articleTitle |
An electrochemiluminescent aptamer switch for a high-throughput assay of an RNA editing reaction.
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pubmed:affiliation |
Department of Pharmacology, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, Non-P.H.S.,
Research Support, Non-U.S. Gov't
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