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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Detalles Bibliográficos
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
Descripción
Sumario: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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