State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, Research Center for Analytical Sciences, College of Chemistry, and School of Medicine, Nankai University, Tianjin, 300071, P. R. China.
Beijing Institutes of Life Science, Chinese Academy of Science, Beijing, 100101, China.
Angew Chem Int Ed Engl. 2023 Jun 19;62(25):e202217230. doi: 10.1002/anie.202217230. Epub 2023 May 9.
Real-time visualization of individual viral mRNA translation activities in live cells is essential to obtain critical details of viral mRNA dynamics and to detect its transient responses to environmental stress. Fluorogenic RNA aptamers are powerful tools for real-time imaging of mRNA in live cells, but monitoring the translation activity of individual mRNAs remains a challenge due to their intrinsic photophysical properties. Here, we develop a genetically encoded turn-on 3,5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI)-binding RNA nanozipper with superior brightness and high photostability by in situ self-assembly of multiple nanozippers along single mRNAs. The nanozipper enables real-time imaging of the mobility and dynamic translation of individual viral mRNAs in live cells, providing information on the spatial dynamics and translational elongation rate of viral mRNAs.
实时可视化活细胞中单个病毒 mRNA 翻译活性对于获得病毒 mRNA 动态的关键细节以及检测其对环境胁迫的瞬时反应至关重要。荧光 RNA 适体是实时成像活细胞中 mRNA 的有力工具，但由于其内在的光物理性质，监测单个 mRNA 的翻译活性仍然是一个挑战。在这里，我们通过多个纳米拉链沿单个 mRNA 原位自组装，开发了一种具有优越亮度和高光稳定性的遗传编码的 3,5-二氟-4-羟基苯亚甲基咪唑啉酮（DFHBI）结合 RNA 纳米拉链。该纳米拉链能够实时成像活细胞中单个病毒 mRNA 的迁移和动态翻译，提供病毒 mRNA 空间动力学和翻译延伸率的信息。
