Wu Yuting, Kong Wentao, Van Stappen Jacqueline, Kong Linggen, Huang Zhimei, Yang Zhenglin, Kuo Yu-An, Chen Yuan-I, He Yujie, Yeh Hsin-Chih, Lu Ting, Lu Yi
Department of Chemistry, The University of Texas at Austin, Austin, Texas 78712, United States.
Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, United States.
J Am Chem Soc. 2025 Jan 15;147(2):1529-1541. doi: 10.1021/jacs.4c09855. Epub 2024 Dec 31.
Genetically encoded fluorescent protein and fluorogenic RNA sensors are indispensable tools for imaging biomolecules in cells. To expand the toolboxes and improve the generalizability and stability of this type of sensor, we report herein a genetically encoded fluorogenic DNA aptamer (GEFDA) sensor by linking a fluorogenic DNA aptamer for dimethylindole red with an ATP aptamer. The design enhances red fluorescence by 4-fold at 650 nm in the presence of ATP. Additionally, upon dimerization, it improves the signal-to-noise ratio by 2-3 folds. We further integrated the design into a plasmid to create a GEFDA sensor for sensing ATP in live bacterial and mammalian cells. This work expanded genetically encoded sensors by employing fluorogenic DNA aptamers, which offer enhanced stability over fluorogenic proteins and RNAs, providing a novel tool for real-time monitoring of an even broader range of small molecular metabolites in biological systems.
基因编码的荧光蛋白和荧光RNA传感器是用于细胞内生物分子成像的不可或缺的工具。为了扩展此类传感器的工具箱并提高其通用性和稳定性，我们在此报告一种基因编码的荧光DNA适配体（GEFDA）传感器，它通过将用于二甲基吲哚红的荧光DNA适配体与ATP适配体连接而成。该设计在ATP存在下，在650nm处可将红色荧光增强4倍。此外，二聚化后，它可将信噪比提高2至3倍。我们进一步将该设计整合到质粒中，以创建用于在活细菌和哺乳动物细胞中检测ATP的GEFDA传感器。这项工作通过使用荧光DNA适配体扩展了基因编码传感器，与荧光蛋白和RNA相比，荧光DNA适配体具有更高的稳定性，为实时监测生物系统中更广泛的小分子代谢物提供了一种新工具。
