Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 Systems
Liang-Zhong Yang, Yang Wang, Si-Qi Li, Run-Wen Yao, Peng-Fei Luan, Huang Wu, Gordon G. Carmichael, Ling-Ling Chen
Journal：MOLECULAR CELL
IF：14.55
DOI：10.1016/j.molcel.2019.10.024
PMID：31757757
Published：2019-11-19
research field：分子生物学细胞生物学基因组学
Abstract
Summary Visualizing the location and dynamics of RNAs in live cells is key to understanding their function. Here, we identify two endonuclease-deficient, single-component programmable RNA-guided and RNA-targeting Cas13 RNases (dCas13s) that allow robust real-time imaging and tracking of RNAs in live cells, even when using single 20- to 27-nt-long guide RNAs. Compared to the aptamer-based MS2-MCP strategy, an optimized dCas13 system is user friendly, does not require genetic manipulation, and achieves comparable RNA-labeling efficiency. We demonstrate that the dCas13 system is capable of labeling NEAT1 , SatIII , MUC4 , and GCN4 RNAs and allows the study of paraspeckle-associated NEAT1 dynamics. Applying orthogonal dCas13 proteins or combining dCas13 and MS2-MCP allows dual-color imaging of RNAs in single cells. Further combination of dCas13 and dCas9 systems allows simultaneous visualization of genomic DNA and RNA transcripts in living cells.
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