Reporter-recruiting bifunctional aptasensor for bioluminescent analytical assays : научное издание

Описание

Тип публикации: статья из журнала

Год издания: 2020

Идентификатор DOI: 10.1039/d0ra05117a

Ключевые слова: the work was supported by the russian science foundation (grant # 16-14-10296), russian state funded budget projects # aaaa-a17-117020210021-7 to icbfm sb ras and # aaaa-a19-119031890015-0 to ibp sb ras.

Аннотация: 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 bioluminesceПоказать полностьюnt 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. 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. © The Royal Society of Chemistry 2020.

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Издание

Журнал: RSC ADVANCES

Выпуск журнала: Vol. 10, Is. 54

Номера страниц: 32393-32399

ISSN журнала: 20462069

Место издания: CAMBRIDGE

Издатель: ROYAL SOC CHEMISTRY

Авторы

  • Davydova Anna (SB RAS, Inst Chem Biol & Fundamental Med, Novosibirsk 630090, Russia)
  • Krasitskaya Vasilisa (SB RAS, Inst Biophys, Fed Res Ctr, Krasnoyarsk Sci Ctr, Krasnoyarsk 660036, Russia)
  • Vorobjev Pavel (SB RAS, Inst Chem Biol & Fundamental Med, Novosibirsk 630090, Russia; Novosibirsk State Univ, Pimgova St 2, Novosibirsk 630090, Russia)
  • Timoshenko Valentina (SB RAS, Inst Chem Biol & Fundamental Med, Novosibirsk 630090, Russia)
  • Tupikin Alexey (SB RAS, Inst Chem Biol & Fundamental Med, Novosibirsk 630090, Russia)
  • Kabilov Marsel (SB RAS, Inst Chem Biol & Fundamental Med, Novosibirsk 630090, Russia)
  • Frank Ludmila (SB RAS, Inst Biophys, Fed Res Ctr, Krasnoyarsk Sci Ctr, Krasnoyarsk 660036, Russia; Siberian Fed Univ, Krasnoyarsk 660041, Russia)
  • Venyaminova Alya (SB RAS, Inst Chem Biol & Fundamental Med, Novosibirsk 630090, Russia)
  • Vorobyeva Mariya (SB RAS, Inst Chem Biol & Fundamental Med, Novosibirsk 630090, Russia)

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