A fluorogenic aptamer can specifically interact with a fluorophore to activate its fluorescence. These nucleic acid-based fluorogenic modules have been dramatically developed over the past decade, and have been used as versatile reporters in the sensor development and for intracellular imaging. In this review, we summarize the design principles, applications, and challenges of the first-generation fluorogenic RNA-based sensors. Moreover, we discuss some strategies to develop next-generation biosensors with improved sensitivity, selectivity, quantification property, and eukaryotic robustness. Using genetically encoded catalytic hairpin assembly strategy as an example, we further introduce a standard protocol to design, characterize, and apply these fluorogenic RNA-based sensors for in vitro detection and cellular imaging of target biomolecules. By incorporating natural RNA machineries, nucleic acid nanotechnology, and systematic evolution approaches, next-generation fluorogenic RNA-based devices can be potentially engineered to be widely applied in cell biology and biomedicine.

中文翻译：

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“第二代”基于荧光 RNA 的传感器


荧光适体可以与荧光团特异性相互作用以激活其荧光。这些基于核酸的荧光模块在过去十年中得到了显着发展，并已在传感器开发和细胞内成像中用作多功能报告基因。在这篇综述中，我们总结了第一代基于荧光 RNA 的传感器的设计原理、应用和挑战。此外，我们还讨论了开发具有更高灵敏度、选择性、定量特性和真核稳健性的下一代生物传感器的一些策略。以基因编码催化发夹组装策略为例，我们进一步介绍了一个标准方案来设计、表征和应用这些基于荧光 RNA 的传感器进行目标生物分子的体外检测和细胞成像。通过结合天然 RNA 机器、核酸纳米技术和系统进化方法，下一代基于荧光 RNA 的设备有可能被设计成广泛应用于细胞生物学和生物医学。