Visualizing RNA dynamics in live cells with bright and stable fluorescent RNAs.,Nature Biotechnology

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Visualizing RNA dynamics in live cells with bright and stable fluorescent RNAs.
Nature Biotechnology ( IF 33.1 ) Pub Date : 2019-09-23 , DOI: 10.1038/s41587-019-0249-1
Xianjun Chen _{1,

2,

3} , Dasheng Zhang _{1,

4} , Ni Su _{1,

3} , Bingkun Bao _{1,

4} , Xin Xie _{1,

3} , Fangting Zuo _{1,

3} , Lipeng Yang _{1,

4} , Hui Wang _{1,

3} , Li Jiang _{1,

4} , Qiuning Lin _{1,

4} , Mengyue Fang _{1,

3} , Ningfeng Li _{1,

4} , Xin Hua _{1,

4} , Zhengda Chen _{1,

3} , Chunyan Bao _{1,

4} , Jinjin Xu ₅ , Wenli Du ₅ , Lixin Zhang ₁ , Yuzheng Zhao _{1,

3} , Linyong Zhu _{1,

4} , Joseph Loscalzo ₆ , Yi Yang _{1,

2,

3}

Affiliation

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.

中文翻译：

使用明亮且稳定的荧光 RNA 可视化活细胞中的 RNA 动力学。

荧光 rna (fr) 是结合和激活荧光染料的适体，已被用于对丰富的细胞 RNA 种类进行成像。然而，诸如低亮度和具有不同光谱特征的染料/适体组合的可用性有限等限制限制了这些工具在活哺乳动物细胞和体内的使用。在这里，我们开发了 Peppers，这是一系列单体、明亮且稳定的 FR，具有从青色到红色的广泛发射最大值。Peppers 可以对活细胞中的不同 RNA 种类进行简单而可靠的成像，而对靶 RNA 的转录、定位和翻译的干扰最小。对单个细胞中蛋白质及其信使 RNA 水平的量化表明，翻译受正常酶动力学控制，但具有显着的异质性。我们进一步表明，Peppers 可用于通过 CRISPR 显示对基因组位点进行成像，用于实时跟踪蛋白质-RNA 束缚，以及用于超分辨率成像。我们相信这些 FR 将成为细胞 RNA 实时成像的有用工具。

更新日期：2019-09-23

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