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A Capture-SELEX Strategy for Multiplexed Selection of RNA Aptamers Against Small Molecules
- Source :
- Methods in Molecular Biology ISBN: 9781493972944
- Publication Year :
- 2018
- Publisher :
- Springer New York, 2018.
-
Abstract
- In vitro selection of aptamers that recognize small organic molecules has proven difficult, in part due to the challenge of immobilizing small molecules on solid supports for SELEX (Systematic Evolution of Ligands by Exponential Enrichment). This study describes the implementation of RNA Capture-SELEX, a selection strategy that uses an RNA library to yield ligand-responsive RNA aptamers targeting small organic molecules in solution. To demonstrate the power of this method we selected several aptamers with specificity towards either the natural sweetener rebaudioside A or the food-coloring agent carminic acid. In addition, Bio-layer interferometry is used to screen clonal libraries of aptamer candidates and is used to interrogate aptamer affinity. The RNA-based Capture-SELEX strategy described here simplifies selection of RNA aptamers against small molecules by avoiding ligand immobilization, while also allowing selection against multiple candidate targets in a single experiment. This makes RNA Capture-SELEX particularly attractive for accelerated development of RNA aptamers targeting small metabolites for incorporation into synthetic riboswitches and for analytical biosensors.
- Subjects :
- 0301 basic medicine
Riboswitch
Bio-layer interferometry
010405 organic chemistry
Chemistry
Aptamer
RNA
Computational biology
01 natural sciences
Small molecule
0104 chemical sciences
03 medical and health sciences
030104 developmental biology
SELEX Aptamer Technique
Biosensor
Systematic evolution of ligands by exponential enrichment
Subjects
Details
- ISBN :
- 978-1-4939-7294-4
- ISBNs :
- 9781493972944
- Database :
- OpenAIRE
- Journal :
- Methods in Molecular Biology ISBN: 9781493972944
- Accession number :
- edsair.doi...........9d24c1dee38a6e380cf7b085dbbc5755