文献类型：期刊文献

英文题名：Visualizing RNA dynamics in live cells with bright and stable fluorescent RNAs

作者：Chen, Xianjun[1,3,4];Zhang, Dasheng[1,2];Su, Ni[1,4];Bao, Bingkun[1,2];Xie, Xin[1,4];Zuo, Fangting[1,4];Yang, Lipeng[1,2];Wang, Hui[1,4];Jiang, Li[1,2];Lin, Qiuning[1,2];Fang, Mengyue[1,4];Li, Ningfeng[1,2];Hua, Xin[1,2];Chen, Zhengda[1,4];Bao, Chunyan[1,2];Xu, Jinjin[5];Du, Wenli[5];Zhang, Lixin[1];Zhao, Yuzheng[1,4];Zhu, Linyong[1,2];Loscalzo, Joseph[6];Yang, Yi[1,3,4]

机构：[1]East China Univ Sci & Technol, Optogenet & Synthet Biol Interdisciplinary Res Ct, State Key Lab Bioreactor Engn, Shanghai Collaborat Innovat Ctr Biomfg Technol, Shanghai, Peoples R China;[2]East China Univ Sci & Technol, Sch Chem & Mol Engn, Shanghai, Peoples R China;[3]Chinese Acad Sci, CAS Ctr Excellence Brain Sci & Intelligence Techn, Inst Neurosci, Shanghai, Peoples R China;[4]East China Univ Sci & Technol, Sch Pharm, Shanghai, Peoples R China;[5]East China Univ Sci & Technol, Sch Informat Sci & Engn, Key Lab Adv Control & Optimizat Chem Processed, Minist Educ, Shanghai, Peoples R China;[6]Harvard Med Sch, Brigham & Womens Hosp, Dept Med, Boston, MA 02115 USA

年份：2019

卷号：37

期号：11

起止页码：1287

外文期刊名：NATURE BIOTECHNOLOGY

收录：;EI(收录号：20194407598624);WOS:【SCI-EXPANDED(收录号:WOS:000494984400016)】；

基金：We thank S.R. Jaffrey at Cornell University for sharing the pAVU6+27-F30-2xdBroccoli construct, Y. Sun at Peking University for the dCas9-GFP construct, C.L. Tucker at Duke University for CIBN/CRY2 constructs. We thank Y. Tang, Y. Liang, L. Chen, Y. Yu and R. Yao for technical assistance and S. Tribuna for secretarial assistance. This research was supported by the National Key Research and Development Program of China (grant no. 2017YFA050400 to Y.Y.), NSFC (grant nos. 21425311 and 21877037 to L. Zhu.; nos. 91857202, 21937004, 31225008 and 31470833 to Y.Y. and no. 31600688 to X.C.), the Shanghai Science and Technology Commission (grant nos. 18JC1411900 and 16430723100 to Y.Y.), the Research Program of State Key Laboratory of Bioreactor Engineering (to Y.Y.) and the China Postdoctoral Science Foundation (grant nos. 2016M59027 and 2017T00277 to X.C.). J.L. was supported by US NIH grant nos. HL61795, HG007690 and GM107618, and by an American Heart Association grant no. D007382.

语种：英文

外文关键词：Cells - Enzyme kinetics - Fluorescence - Mammals - Cytology

摘要：Fluorescent RNAs (FRs), aptamers that bind and activate fluorescent dyes, have been used to image abundant cellular RNA species. However, limitations such as low brightness and limited availability of dye/aptamer combinations with different spectral characteristics have limited use of these tools in live mammalian cells and in vivo. Here, we develop Peppers, a series of monomeric, bright and stable FRs with a broad range of emission maxima spanning from cyan to red. Peppers allow simple and robust imaging of diverse RNA species in live cells with minimal perturbation of the target RNA's transcription, localization and translation. Quantification of the levels of proteins and their messenger RNAs in single cells suggests that translation is governed by normal enzyme kinetics but with marked heterogeneity. We further show that Peppers can be used for imaging genomic loci with CRISPR display, for real-time tracking of protein-RNA tethering, and for super-resolution imaging. We believe these FRs will be useful tools for live imaging of cellular RNAs.