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 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.
Издание
Журнал: RSC ADVANCES
Выпуск журнала: Vol. 10, Is. 54
Номера страниц: 32393-32399
ISSN журнала: 20462069
Место издания: CAMBRIDGE
Издатель: ROYAL SOC CHEMISTRY
Персоны
- 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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