A Post-Labeling Approach for the Characterization and Quantification of RNA Modifications Based on Site-Directed Cleavage by DNAzymes
- Publication type:
- Journal article
- Metadata:
-
- Autoren
- Madeleine Meusburger
- Martin Hengesbach
- Mark Helm
- DOI
- 10.1007/978-1-61779-018-8_16
- eISSN
- 1940-6029
- ISSN
- 1064-3745
- Online publication date
- 2011
- Paginierung
- 259 - 270
- Datum der Veröffentlichung
- 2011
- Status
- Published
- Herausgeber
- Humana Press
- Herausgeber URL
- http://dx.doi.org/10.1007/978-1-61779-018-8_16
- Datum der Datenerfassung
- 2023
- Titel
- A Post-Labeling Approach for the Characterization and Quantification of RNA Modifications Based on Site-Directed Cleavage by DNAzymes
Data source: Crossref
- Other metadata sources:
-
- Abstract
- Deoxyribozymes or DNAzymes are small DNA molecules with catalytic activity originating from in vitro selection experiments. Variants of the two most popular DNAzymes with RNase activity, the 10-23 DNAzyme and the 8-17 DNAzyme, promote efficient in vitro cleavage of the phosphodiester bond in at least 11 out of 16 possible dinucleotide permutations. Judicious choice of the sequences flanking the active core of the DNAzymes permits to direct cleavage activity with high sequence specificity. Here, the harnessing of these features for the analysis of RNA nucleotide modifications by a post-labeling approach is described in detail. DNAzymes are designed such that RNase cleavage is directed precisely to the 5' end of the nucleotide to be analyzed. Iterative complex formation of DNAzyme and RNA substrate and subsequent cleavage are performed by temperature cycling. The DNAzyme activity liberates the analyte nucleotide on the very 5'-end of an RNA fragment, whose hydroxyl group can be conveniently phosphorylated with (32)P. The labeled RNA is digested to mononucleotides, and analyzed by thin layer chromatography.
- Addresses
- Institute of Pharmacy and Molecular Biotechnology, University of Heidelberg, Heidelberg, Germany.
- Autoren
- Madeleine Meusburger
- Martin Hengesbach
- Mark Helm
- DOI
- 10.1007/978-1-61779-018-8_16
- eISSN
- 1940-6029
- Externe Identifier
- PubMed Identifier: 21370054
- Open access
- false
- ISSN
- 1064-3745
- Zeitschrift
- Methods in molecular biology (Clifton, N.J.)
- Schlüsselwörter
- Humans
- DNA, Catalytic
- Nucleotides
- RNA
- Chromatography, Thin Layer
- Base Sequence
- Sprache
- eng
- Medium
- Paginierung
- 259 - 270
- Datum der Veröffentlichung
- 2011
- Status
- Published
- Datum der Datenerfassung
- 2011
- Titel
- A post-labeling approach for the characterization and quantification of RNA modifications based on site-directed cleavage by DNAzymes.
- Sub types
- Research Support, Non-U.S. Gov't
- Journal Article
- Ausgabe der Zeitschrift
- 718
Data source: Europe PubMed Central
- Abstract
- Deoxyribozymes or DNAzymes are small DNA molecules with catalytic activity originating from in vitro selection experiments. Variants of the two most popular DNAzymes with RNase activity, the 10-23 DNAzyme and the 8-17 DNAzyme, promote efficient in vitro cleavage of the phosphodiester bond in at least 11 out of 16 possible dinucleotide permutations. Judicious choice of the sequences flanking the active core of the DNAzymes permits to direct cleavage activity with high sequence specificity. Here, the harnessing of these features for the analysis of RNA nucleotide modifications by a post-labeling approach is described in detail. DNAzymes are designed such that RNase cleavage is directed precisely to the 5' end of the nucleotide to be analyzed. Iterative complex formation of DNAzyme and RNA substrate and subsequent cleavage are performed by temperature cycling. The DNAzyme activity liberates the analyte nucleotide on the very 5'-end of an RNA fragment, whose hydroxyl group can be conveniently phosphorylated with (32)P. The labeled RNA is digested to mononucleotides, and analyzed by thin layer chromatography.
- Autoren
- Madeleine Meusburger
- Martin Hengesbach
- Mark Helm
- Autoren-URL
- https://www.ncbi.nlm.nih.gov/pubmed/21370054
- DOI
- 10.1007/978-1-61779-018-8_16
- eISSN
- 1940-6029
- Zeitschrift
- Methods Mol Biol
- Schlüsselwörter
- Base Sequence
- Chromatography, Thin Layer
- DNA, Catalytic
- Humans
- Nucleotides
- RNA
- Sprache
- eng
- Country
- United States
- Paginierung
- 259 - 270
- Datum der Veröffentlichung
- 2011
- Status
- Published
- Datum, an dem der Datensatz öffentlich gemacht wurde
- 2011
- Titel
- A post-labeling approach for the characterization and quantification of RNA modifications based on site-directed cleavage by DNAzymes.
- Sub types
- Journal Article
- Research Support, Non-U.S. Gov't
- Ausgabe der Zeitschrift
- 718
Data source: PubMed
- Beziehungen:
- Property of