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