Subcellular localizations of RNAs can be imaged in vivo with genetically encoded reporters consisting of a sequence-specific RNA-binding protein (RBP) fused to a fluorescent protein. Several such reporter systems have been described based on RBPs that recognize RNA stem-loops. Here we describe RNA tagging for imaging with an inactive mutant of the bacterial endonuclease Csy4, which has a significantly higher affinity for its cognate stem-loop than alternative systems. This property allows for sensitive imaging with only few tandem copies of the target stem-loop inserted into the RNA of interest.
CITATION STYLE
Burnett, D., Lichius, A., & Tilsner, J. (2020). RNA imaging with rnase-inactivated Csy4 in plants and filamentous fungi. In Methods in Molecular Biology (Vol. 2166, pp. 157–178). Humana Press Inc. https://doi.org/10.1007/978-1-0716-0712-1_9
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