Spinach-based RNA mimicking GFP in plant cells

Abstract

Spinach RNA-mimicking GFP (S-RMG) has been successfully used to monitor cellular RNAs including microRNAs in bacterium, yeast, and human cells. However, S-RMG has not been established in plants. In this study, we found that like bacterial, yeast, and human cellular tRNAs, plant tRNAs such as tRNALys can protect and/or stabilize the Spinach RNA aptamer interaction with the fluorophore DFHBI enabling detectable levels of green fluorescence to be emitted. The tRNALys-Spinach-tRNALys, once delivered into “chloroplast-free” onion epidermal cells can emit strong green fluorescence in the presence of DFHBI. Our results demonstrate for the first time that Spinach-based RNA visualization has the potential for in vivo monitoring of RNAs in plant cells.

Traveling with purpose: cell-to-cell transport of plant mRNAs

“…However, to observe the cellto-cell trafficking of mRNAs, a new reporter system that does not affect movement is needed. For example, the spinach RNA-fluorophore complex is a smaller tag that resembles green fluorescent protein (GFP) and has been applied to plants [98], and an RNA labeling system using a bacterial RBP has been used to visualize the cell-to-cell movement of MP RNA [99].…”

Aggregation-Induced Emission Luminogens: A New Possibility for Efficient Visualization of RNA in Plants

“…The combination of DFHBI and 24-2 RNA emits a fluorescent color like that of spinach; thus, it was named Spinach [ 34 ]. The photobleaching effect has caused many obstacles to the application of RNA aptamer-based technology in plants, even though there are only two successfully reported cases [ 35 , 36 ]. Even so, multiple RNA aptamer and fluorophore combinations were still developed, as shown in Table 1 .…”