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Issue 54, 2020, Issue in Progress
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Reporter-recruiting bifunctional aptasensor for bioluminescent analytical assays

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Abstract

We report a novel bioluminescent aptasensor, which consists of 2′-F-RNA aptamer modules joined into a bi-specific aptamer construct. One aptamer module binds the analyte, then after structural rearrangement the second module recruits non-covalently Ca2+-dependent photoprotein obelin from the solution, thus providing a bioluminescent signal. This concept allows using free protein as a reporter, which brings such advantages as no need for aptamer–protein conjugation, a possibility of thermal re-folding of aptamer component with no harm to a protein, and simpler detection protocol. We developed the new 2′-F-RNA aptamer for obelin, and proposed the strategy for engineering structure-switching bi-modular aptamer constructs which bind the analyte and the obelin in a sequential manner. With the use of hemoglobin as a model analyte, we showed the feasibility of utilizing the aptasensor in a fast and straightforward bioluminescent microplate assay. With a proper design of a secondary structure, this strategy of aptasensor engineering might be further extended to bi-specific aptamer-based bioluminescent sensors for other analytes of interest.

Graphical abstract: Reporter-recruiting bifunctional aptasensor for bioluminescent analytical assays

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Supplementary files

Article information


Submitted
10 Jun 2020
Accepted
21 Aug 2020
First published
01 Sep 2020

This article is Open Access

RSC Adv., 2020,10, 32393-32399
Article type
Paper

Reporter-recruiting bifunctional aptasensor for bioluminescent analytical assays

A. Davydova, V. Krasitskaya, P. Vorobjev, V. Timoshenko, A. Tupikin, M. Kabilov, L. Frank, A. Venyaminova and M. Vorobyeva, RSC Adv., 2020, 10, 32393
DOI: 10.1039/D0RA05117A

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