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

Michael Smith, Emmanuelle Querido, Pascal Chartrand, Agnel Sfeir

STAR protocols 2020 Dec 18

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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). © 2020 The Authors.

Citation

Michael Smith, Emmanuelle Querido, Pascal Chartrand, Agnel Sfeir. Quantitative Imaging of MS2-Tagged hTR in Cajal Bodies: Photobleaching and Photoactivation. STAR protocols. 2020 Dec 18;1(3):100112