DNA display of folded RNA libraries enabling RNA-SELEX without reverse transcription
| Autor: | Isaac J. Krauss, J. S. Temme, I. S. MacPherson |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Riboswitch RNA Folding RNA-dependent RNA polymerase Computational biology 010402 general chemistry 01 natural sciences Article Catalysis 03 medical and health sciences Transcription (biology) Materials Chemistry RNA Catalytic Ligase ribozyme Gene Library Transcription bubble Chemistry SELEX Aptamer Technique Metals and Alloys RNA DNA Reverse Transcription General Chemistry Aptamers Nucleotide Molecular biology 0104 chemical sciences Surfaces Coatings and Films Electronic Optical and Magnetic Materials 030104 developmental biology RNA editing Coding strand Ceramics and Composites |
| Zdroj: | Chemical Communications. 53:2878-2881 |
| ISSN: | 1364-548X 1359-7345 |
| DOI: | 10.1039/c6cc09991b |
| Popis: | A method for the physical attachment of folded RNA libraries to their encoding DNA is presented as a way to circumvent the reverse transcription step during systematic evolution of RNA ligands by exponential enrichment (RNA-SELEX). A DNA library is modified with one isodC base to stall T7 polymerase and a 5′ “capture strand” which anneals to the nascent RNA transcript. This method is validated in a selection of RNA aptamers against human α-thrombin with dissociation constants in the low nanomolar range. This method will be useful in the discovery of RNA aptamers and ribozymes containing base modifications that make them resistant to accurate reverse transcription. |
| Databáze: | OpenAIRE |
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