Selection of Aptamers for Metabolic Sensing and Construction of Optical Nanosensors

Yi Long, Franziska Pfeiffer, Günter Mayer, Tine Daa Schrøder, Veli Cengiz Özalp, Lars Folke Olsen

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

Abstract

Optical nanosensors are based on particles with diameters from 20 to 200 nm containing sensory elements. The latter are comprised of one or more signaling molecules and one or more references, which allow measurements to be ratiometric and hence independent on the amount of sensor. The signaling molecules may range from simple ion-binding fluorophores, e.g., pH-sensitive dyes, to complex biochemical assays. Aptamers are ideal for use in nanosensors because they are relatively easy to modify chemically and hence to transform into signaling molecules, and their binding affinities may be fine-tuned to a desired measuring range in the selection process. Here we first describe the selection of metabolite binding aptamers, how they are transformed into signaling molecules using a molecular beacon construct and then how they are inserted into nanoparticles. Finally, we briefly describe how the sensors are calibrated before inserted into cells to measure metabolite concentration in real time. As examples we present aptamers binding to key metabolites in cells: ATP and fructose 1, 6-bisphosphate (FBP).

Original languageEnglish
Title of host publicationNucleic Acid Aptamers : Selection, Characterization, and Application
EditorsGünter Mayer
VolumePart I
PublisherHumana Press
Publication date2016
Pages3-19
Chapter1
ISBN (Print)978-1-4939-3196-5
ISBN (Electronic)978-1-4939-3197-2
DOIs
Publication statusPublished - 2016
SeriesMethods in Molecular Biology
Volume1380
ISSN1064-3745

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