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
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author Wang, Yang
Yang, Liang-Zhong
Chen, Ling-Ling
author_facet Wang, Yang
Yang, Liang-Zhong
Chen, Ling-Ling
author_sort Wang, Yang
collection PubMed
description 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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spelling pubmed-75802062020-10-26 Protocol for Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 System Wang, Yang Yang, Liang-Zhong Chen, Ling-Ling STAR Protoc Protocol 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). Elsevier 2020-06-03 /pmc/articles/PMC7580206/ /pubmed/33111085 http://dx.doi.org/10.1016/j.xpro.2020.100037 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Wang, Yang
Yang, Liang-Zhong
Chen, Ling-Ling
Protocol for Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 System
title Protocol for Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 System
title_full Protocol for Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 System
title_fullStr Protocol for Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 System
title_full_unstemmed Protocol for Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 System
title_short Protocol for Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 System
title_sort protocol for dynamic imaging of rna in living cells by crispr-cas13 system
topic Protocol
url 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
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