文献类型：期刊文献

英文题名：Imaging intracellular metabolite and protein changes in live mammalian cells with bright fluorescent RNA-based genetically encoded sensors

作者：Fang, Mengyue[1,2];Li, Huiwen[1,2];Xie, Xin[1,2];Wang, Hui[1];Jiang, Ying[1,2];Li, Tianyu[1,2];Zhang, Bibi[1,2];Jiang, Xin[1,2];Cao, Yueyang[1,2];Zhang, Rui[1,2];Zhang, Dasheng[1];Zhao, Yuzheng[1,2];Zhu, Linyong[1,3];Chen, Xianjun[1,2];Yang, Yi[1,2]

机构：[1]East China Univ Sci & Technol, Optogenet & Synthet Biol Interdisciplinary Res Ctr, State Key Lab Bioreactor Engn, 130 Mei Long Rd, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Shanghai Frontiers Sci Ctr Optogenet Tech Cell Met, Sch Pharm, 130 Mei Long Rd, Shanghai 200237, Peoples R China;[3]Shanghai Jiao Tong Univ, Sch Biomed Engn, 800 Dong Chuan Rd, Shanghai 200240, Peoples R China

年份：2023

卷号：235

外文期刊名：BIOSENSORS & BIOELECTRONICS

收录：;EI(收录号：20232214170413);WOS:【SCI-EXPANDED(收录号:WOS:001009113100001)】；

基金：We thank Ni Su, Zhengda Chen, Fangting Zuo and Renmei Liu for technical assistance. This research was supported by the National Key Research and Development Program of China (2019YFA0904800 to Y. Y., 2022YFC3400100 to Y.Y. and X.C.), STI2030-Major Projects (2021ZD0202200 and 2021ZD0202203 to X.C.), NSFC (32121005, 32150028, 91857202, and 21937004 to Y.Y., 32250009 and 31600688 to X.C.), the Shanghai Municipal Education Commission-Frontier Science Research Base of Optogenetic Techniques for Cell Metabolism (2021 Sci & Tech 03-28) to Y.Y. and X.C., the Shanghai Rising-Star Program to X.C., the State Key Laboratory of Bioreactor Engineering to Y.Y. and X.C., the Fundamental Research Funds for the Central Universities to Y.Y. and X.C.

语种：英文

外文关键词：Genetically encoded sensor; Fluorogenic RNA aptamer; Pepper; Metabolite and protein dynamics

摘要：Fluorescent RNA (FR)-based genetically encoded sensors have been engineered to detect various essential metabolites in living systems. However, the unfavorable characteristics of FR impede sensor applications. Here, we describe a strategy for converting Pepper fluorescent RNA into a series of fluorescent sensors to detect their cognate targets both in vitro and in live cells. Compared to previously developed FR-based sensors, Pepper-based sensors exhibited expanded emission of up to 620 nm and markedly improved cellular brightness, allowing robust and real-time monitoring of the pharmacologic-triggered dynamics changes in the intracellular level of Sadenosylmethionine (SAM) and the optogenetic manipulated protein translocation in live mammalian cells. Furthermore, signal amplification in fluorescence imaging of the target was achieved using the CRISPR-display strategy by incorporating a Pepper-based sensor into the sgRNA scaffold. Together, these results demonstrate that Pepper can be readily developed into high-performance FR-based sensors to detect various cellular targets.