Visualizing RNA dynamics in live cells with bright and stable fluorescent RNAs
Chen Xianjun, Zhang Dasheng, Su Ni, Bao Bingkun, Xie Xin, Zuo Fangting, Yang Lipeng, Wang Hui, Jiang Li, Lin Qiuning, Fang Mengyue, Li Ningfeng, Hua Xin, Chen Zhengda, Bao Chunyan, Xu Jinjin, Du Wenl
Journal：NATURE BIOTECHNOLOGY
IF：31.86
DOI：10.1038/s41587-019-0249-1
PMID：31548726
Published：2019-09-23
research field：分子生物学细胞生物学生物技术
Abstract
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.
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