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