Reporter-recruiting bifunctional aptasensor for bioluminescent analytical assays

Anna Davydova, Vasilisa Krasitskaya, Pavel Vorobjev, Valentina Timoshenko, Alexey Tupikin, Marsel Kabilov, Ludmila Frank, Alya Venyaminova, Mariya Vorobyeva

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

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.

Original languageEnglish
Pages (from-to)32393-32399
Number of pages7
JournalRSC Advances
Volume10
Issue number54
DOIs
Publication statusPublished - 1 Sep 2020

Keywords

  • DNA APTAMER
  • RNA APTAMER
  • OBELIN
  • PURIFICATION
  • EXPRESSION
  • SEQUENCES
  • CLONING
  • TARGET

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