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

Abstract

Summary: 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).