2. Academic Validation
  3. Visualizing RNA dynamics in live cells with bright and stable fluorescent RNAs

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

  • Nat Biotechnol. 2019 Nov;37(11):1287-1293. 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 6 7,  Joseph Loscalzo 8,  Yi Yang 9 10 11
Affiliations

Affiliations

  • 1 Optogenetics and Synthetic Biology Interdisciplinary Research Center, State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and Technology, Shanghai, China.
  • 2 CAS Center for Excellence in Brain Science and Intelligence Technology, Institute of Neuroscience, Chinese Academy of Sciences, Shanghai, China.
  • 3 School of Pharmacy, East China University of Science and Technology, Shanghai, China.
  • 4 School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai, China.
  • 5 Key Laboratory of Advanced Control and Optimization for Chemical Processed of Ministry of Education, School of information Science and Engineering, East China University of Science and Technology, Shanghai, China.
  • 6 Optogenetics and Synthetic Biology Interdisciplinary Research Center, State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and Technology, Shanghai, China. linyongzhu@ecust.edu.cn.
  • 7 School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai, China. linyongzhu@ecust.edu.cn.
  • 8 Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.
  • 9 Optogenetics and Synthetic Biology Interdisciplinary Research Center, State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and Technology, Shanghai, China. yiyang@ecust.edu.cn.
  • 10 CAS Center for Excellence in Brain Science and Intelligence Technology, Institute of Neuroscience, Chinese Academy of Sciences, Shanghai, China. yiyang@ecust.edu.cn.
  • 11 School of Pharmacy, East China University of Science and Technology, Shanghai, China. yiyang@ecust.edu.cn.
  • # Contributed equally.
PMID: 31548726 DOI: 10.1038/s41587-019-0249-1
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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