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Updated: Dec 3, 2025
Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons
Published on: August 6, 2014
Protocol for Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 System
Yang Wang1, Liang-Zhong Yang1, Ling-Ling Chen1,2
1State Key Laboratory of Molecular Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, University of Chinese Academy of Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China.
Abstract:
This protocol uses endonuclease-dead, programmable RNA-guided RNA-targeting Cas13 RNases (d)Cas13 proteins fused with fluorescent proteins to visualize and track RNA dynamics in live cells. This protocol details several aspects of the procedure, including gRNA design, fluorescent protein selection, nuclear localization signal adjustment, raw data analysis, operation steps, and extended optional applications that have been successfully applied in the visualization of NEAT1, SatIII, MUC4, and GCN4 RNAs. For complete information on the use and execution of this protocol, please refer to Yang et al. (2019).
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