华东理工大学杨弋团队与朱麟勇团队近日合作研发出可用于活细胞RNA动态成像的荧光RNA。该研究2019年9月23日在线发表于《自然—生物技术》。

研究人员开发了一系列单体、明亮且稳定的FR，叫做Peppers，其最大发射范围从青色到红色。Peppers可以对活细胞中的各种RNA进行简单而强大的成像，而对目标RNA的转录、定位和翻译的干扰却最小。定量分析单个细胞中蛋白质及其信使RNA的水平表明，翻译受正常的酶动力学控制，但具有明显的异质性。

研究人员进一步证明，Peppers可用于通过CRISPR展示对基因组基因座进行成像，实时跟踪蛋白质-RNA拴链以及进行超分辨率成像。研究人员相信这些FR将是细胞RNA实时成像的有用工具。

研究人员表示，荧光RNA（FR），即结合并激活荧光染料的核酸适配体，已用于丰富的细胞内RNA成像。然而，诸如低亮度和具有不同光谱特性的染料/适配体组合的有限可用性限制了这些工具在活的哺乳动物细胞和体内的使用。

附：英文原文

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

Author: Xianjun Chen, Dasheng Zhang, Ni Su, Bingkun Bao, Xin Xie, Fangting Zuo, Lipeng Yang, Hui Wang, Li Jiang, Qiuning Lin, Mengyue Fang, Ningfeng Li, Xin Hua, Zhengda Chen, Chunyan Bao, Jinjin Xu, Wenli Du, Lixin Zhang, Yuzheng Zhao, Linyong Zhu, Joseph Loscalzo, Yi Yang

Issue&Volume: 2019-09-23

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.

DOI: 10.1038/s41587-019-0249-1

Source:https://www.nature.com/articles/s41587-019-0249-1