Department of Chemistry, University of Massachusetts, Amherst, Massachusetts 01003, United States.
School of Chemistry and Chemical Engineering, Nanjing University of Science and Technology, Nanjing 210094, China.
ACS Sens. 2023 Jan 27;8(1):308-316. doi: 10.1021/acssensors.2c02213. Epub 2023 Jan 6.
RNA-based nanostructures and molecular devices have become popular for developing biosensors and genetic regulators. These programmable RNA nanodevices can be genetically encoded and modularly engineered to detect various cellular targets and then induce output signals, most often a fluorescence readout. Although powerful, the high reliance of fluorescence on the external excitation light raises concerns about its high background, photobleaching, and phototoxicity. Bioluminescence signals can be an ideal complementary readout for these genetically encoded RNA nanodevices. However, RNA-based real-time bioluminescent reporters have been rarely developed. In this study, we reported the first type of genetically encoded RNA-based bioluminescence resonance energy transfer (BRET) sensors that can be used for real-time target detection in living cells. By coupling a luciferase bioluminescence donor with a fluorogenic RNA-based acceptor, our BRET system can be modularly designed to image and detect various cellular analytes. We expect that this novel RNA-based bioluminescent system can be potentially used broadly in bioanalysis and nanomedicine for engineering biosensors, characterizing cellular RNA-protein interactions, and high-throughput screening or imaging.
基于 RNA 的纳米结构和分子器件已成为开发生物传感器和遗传调节剂的热门选择。这些可编程 RNA 纳米器件可以通过遗传编码和模块化设计来检测各种细胞靶标，然后诱导输出信号，通常是荧光读数。尽管功能强大，但荧光对外部激发光的高度依赖引发了人们对其高背景、光漂白和光毒性的担忧。生物发光信号可以成为这些遗传编码 RNA 纳米器件的理想互补读数。然而，基于 RNA 的实时生物发光报告器很少得到开发。在本研究中，我们报告了第一种基于 RNA 的遗传编码生物发光共振能量转移 (BRET) 传感器，可用于活细胞中的实时目标检测。通过将荧光素酶生物发光供体与荧光 RNA 基受体偶联，我们的 BRET 系统可以进行模块化设计，以成像和检测各种细胞分析物。我们预计，这种新型基于 RNA 的生物发光系统有望在生物分析和纳米医学中广泛用于工程生物传感器、表征细胞 RNA-蛋白质相互作用以及高通量筛选或成像。
