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Genomic DNA i-motifs as fast sensors responsive to near-physiological pH microchangesстатья
Статья опубликована в высокорейтинговом журнале
Информация о цитировании статьи получена из
Web of Science,
Scopus
Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 17 февраля 2021 г.
- Авторы: Turaev Anton V., Isaakova Ekaterina A., Severov Vjacheslav V., Bogomazova Alexandra N., Zatsepin Timofei S., Sardushkin Makar V., Aralov Andrey V., Lagarkova Maria A., Pozmogova Galina E., Varizhuk Anna M.
- Журнал: Biosensors and Bioelectronics
- Год издания: 2020
- Издательство: Pergamon Press Ltd.
- Местоположение издательства: United Kingdom
- Номер статьи: 112864
- DOI: 10.1016/j.bios.2020.112864
- Аннотация: We report the design of robust sensors for measuring intracellular pH, based on the native DNA i-motifs (iMs) found in neurodegeneration- or carcinogenesis-related genes. Those iMs appear to be genomic regulatory elements and might modulate transcription in response to pH stimuli. Given their intrinsic sensitivity to minor pH changes within the physiological range, such noncanonical DNA structures can be used as sensor core elements without additional modules other than fluorescent labels or quenchers. We focused on several iMs that exhibited fast folding/unfolding kinetics. Using stopped-flow techniques and FRET-melting/annealing assays, we confirmed that the rates of temperature-driven iM-ssDNA transitions correlate with the rates of the pH-driven transitions. Thus, we propose FRET-based hysteresis analysis as an express method for selecting sensors with desired kinetic characteristics. For the leading fast-response sensor, we optimized the labelling scheme and performed intracellular calibration. Unlike the commonly used small-molecule pH indicators, that sensor was transferred efficiently to cell nuclei. Considering its favourable kinetic characteristics, the sensor can be used for monitoring proton dynamics in the nucleus. These results argue that the ‘genome-inspired’ design is a productive approach to the development of biocompatible molecular tools.
- Добавил в систему: Зацепин Тимофей Сергеевич