Protocol for Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 System

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,...

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Detalles Bibliográficos
Autores principales: Wang, Yang, Yang, Liang-Zhong, Chen, Ling-Ling
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7580206/
https://www.ncbi.nlm.nih.gov/pubmed/33111085
http://dx.doi.org/10.1016/j.xpro.2020.100037
Descripción
Sumario: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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