Quantitative Imaging of MS2-Tagged hTR in Cajal Bodies: Photobleaching and Photoactivation

Advances in imaging technologies, gene editing, and fluorescent molecule development have made real-time imaging of nucleic acids practical. Here, we detail methods for imaging the human telomerase RNA template, hTR via the use of three inserted MS2 stem loops and cognate MS2 coat protein (MCP) tagg...

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Autores principales: Smith, Michael, Querido, Emmanuelle, Chartrand, Pascal, Sfeir, Agnel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7756913/
https://www.ncbi.nlm.nih.gov/pubmed/33377008
http://dx.doi.org/10.1016/j.xpro.2020.100112
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author Smith, Michael
Querido, Emmanuelle
Chartrand, Pascal
Sfeir, Agnel
author_facet Smith, Michael
Querido, Emmanuelle
Chartrand, Pascal
Sfeir, Agnel
author_sort Smith, Michael
collection PubMed
description Advances in imaging technologies, gene editing, and fluorescent molecule development have made real-time imaging of nucleic acids practical. Here, we detail methods for imaging the human telomerase RNA template, hTR via the use of three inserted MS2 stem loops and cognate MS2 coat protein (MCP) tagged with superfolder GFP or photoactivatable GFP. These technologies enable tracking of the dynamics of RNA species through Cajal bodies and offer insight into their residence time in Cajal bodies through photobleaching and photoactivation experiments. For complete details on the use and execution of this protocol, please refer to Laprade et al. (2020).
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spelling pubmed-77569132020-12-28 Quantitative Imaging of MS2-Tagged hTR in Cajal Bodies: Photobleaching and Photoactivation Smith, Michael Querido, Emmanuelle Chartrand, Pascal Sfeir, Agnel STAR Protoc Protocol Advances in imaging technologies, gene editing, and fluorescent molecule development have made real-time imaging of nucleic acids practical. Here, we detail methods for imaging the human telomerase RNA template, hTR via the use of three inserted MS2 stem loops and cognate MS2 coat protein (MCP) tagged with superfolder GFP or photoactivatable GFP. These technologies enable tracking of the dynamics of RNA species through Cajal bodies and offer insight into their residence time in Cajal bodies through photobleaching and photoactivation experiments. For complete details on the use and execution of this protocol, please refer to Laprade et al. (2020). Elsevier 2020-09-24 /pmc/articles/PMC7756913/ /pubmed/33377008 http://dx.doi.org/10.1016/j.xpro.2020.100112 Text en © 2020 The Authors 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
Smith, Michael
Querido, Emmanuelle
Chartrand, Pascal
Sfeir, Agnel
Quantitative Imaging of MS2-Tagged hTR in Cajal Bodies: Photobleaching and Photoactivation
title Quantitative Imaging of MS2-Tagged hTR in Cajal Bodies: Photobleaching and Photoactivation
title_full Quantitative Imaging of MS2-Tagged hTR in Cajal Bodies: Photobleaching and Photoactivation
title_fullStr Quantitative Imaging of MS2-Tagged hTR in Cajal Bodies: Photobleaching and Photoactivation
title_full_unstemmed Quantitative Imaging of MS2-Tagged hTR in Cajal Bodies: Photobleaching and Photoactivation
title_short Quantitative Imaging of MS2-Tagged hTR in Cajal Bodies: Photobleaching and Photoactivation
title_sort quantitative imaging of ms2-tagged htr in cajal bodies: photobleaching and photoactivation
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7756913/
https://www.ncbi.nlm.nih.gov/pubmed/33377008
http://dx.doi.org/10.1016/j.xpro.2020.100112
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AT chartrandpascal quantitativeimagingofms2taggedhtrincajalbodiesphotobleachingandphotoactivation
AT sfeiragnel quantitativeimagingofms2taggedhtrincajalbodiesphotobleachingandphotoactivation

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