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λN-GFP: an RNA reporter system for live-cell imaging

Abstract

We describe a GFP-based RNA reporter system (λN-GFP) to visualize RNA molecules in live mammalian cells. It consists of GFP fused to an arginine-rich peptide derived from the phage λ N protein, λN22, which binds a unique minimal RNA motif and can be used to tag any RNA molecule. λN-GFP uses a small and easy to engineer RNA tag, reducing the likelihood of perturbing the function of the tagged RNA molecule.

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Figure 1: Schematic representation of the GFP-based RNA reporter system.
Figure 2: 4 × λN22-3 × mEGFP-M9 is exported out of the nucleus only in the presence of boxB-tagged mRNA (a–d).
Figure 3: 4 × λN22-3 × mEGFP-M9 serves as a reporter of mRNA localization in the cytoplasm.

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Acknowledgements

N.D. thanks J. Beaudouin, P. Lénárt and Y. Trottier for lively discussions.

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Correspondence to Jan Ellenberg.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–2, Supplementary Methods (PDF 432 kb)

Supplementary Movie 1

Time-lapse recording of a NRK cell expressing the tagged mRNA mRFP-4boxB-zipcode and the translated protein mRFP. (MOV 2517 kb)

Supplementary Movie 2

Time-lapse recording of post-mitotic NRK cells expressing the tagged mRNA Srprb-mRFP-16boxB and the translated protein Srprb-mRFP. (MOV 1972 kb)

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Daigle, N., Ellenberg, J. λN-GFP: an RNA reporter system for live-cell imaging. Nat Methods 4, 633–636 (2007). https://doi.org/10.1038/nmeth1065

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  • DOI: https://doi.org/10.1038/nmeth1065

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