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Live-cell imaging of single mRNA dynamics using split superfolder green fluorescent proteins with minimal background

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Title
Live-cell imaging of single mRNA dynamics using split superfolder green fluorescent proteins with minimal background
Author(s)
Park S.Y.; Moon H.C.; Park H.Y.
Publication Date
2020-01
Journal
RNA-A PUBLICATION OF THE RNA SOCIETY, v.26, no.1, pp.101 - 109
Publisher
COLD SPRING HARBOR LAB PRESS
Abstract
© 2020 Park et al.The MS2 system, with an MS2 binding site (MBS) and an MS2 coat protein fused to a fluorescent protein (MCP-FP), has been widely used to fluorescently label mRNA in live cells. However, one of its limitations is the constant background fluorescence signal generated from freeMCP-FPs. To overcome this obstacle, we used a superfolder GFP (sfGFP) split into two or three nonfluorescent fragments that reassemble and emit fluorescence only when bound to the target mRNA. Using the high-affinity interactions of bacteriophage coat proteins with their corresponding RNA binding motifs, we showed that the nonfluorescent sfGFP fragments were successfully brought close to each other to reconstitute a complete sfGFP. Furthermore, real-time mRNA dynamics inside the nucleus as well as the cytoplasm were observed by using the split sfGFPs with the MS2-PP7 hybrid system. Our results demonstrate that the split sfGFP systems are useful tools for background- free imaging of mRNA with high spatiotemporal resolution
URI
https://pr.ibs.re.kr/handle/8788114/7034
ISSN
1355-8382
Appears in Collections:
Center for RNA Research(RNA 연구단) > Journal Papers (저널논문)
Files in This Item:
RNA-2019-Park-rna.067835.118.pdfDownload

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