Visualizing RNA dynamics in live cells with bright and stable fluorescent RNAs

被引：254

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

来源：

NATURE BIOTECHNOLOGY | 2019年 / 37卷 / 11期

基金：

中国博士后科学基金;

关键词：

FLUOROGENIC CYANINE DYES; GENE-EXPRESSION; TRANSLATION DYNAMICS; FLUOROPHORE-BINDING; STRUCTURAL BASIS; APTAMERS; PROTEIN; MIMICS; QUANTIFICATION; LOCALIZATION;

D O I：

10.1038/s41587-019-0249-1

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

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.

引用

页码：1287 / +

页数：10

共 60 条

[1] Chromatin domains and the interchromatin compartment form structurally defined and functionally interacting nuclear networks [J].