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Imaging bacterial protein expression using genetically encoded RNA sensors

Abstract

The difficulties in imaging the dynamics of protein expression in live bacterial cells can be overcome by using fluorescent sensors based on Spinach, an RNA that activates the fluorescence of a small-molecule fluorophore. These RNAs selectively bind target proteins and exhibit fluorescence increases that enable protein expression to be imaged in living Escherichia coli. These sensors are key components of a generalizable strategy to image protein expression in a single bacterium in real time.

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Figure 1: Sensitive and specific detection of proteins in vitro using Spinach-based sensors.
Figure 2: Visualization of MS2 coat protein (MCP) synthesis in individual cells after MS2 phage infection.

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Acknowledgements

We thank J.S. Paige for useful comments and suggestions and F. Dardel (Université Paris Descartes) for providing plasmids containing the tRNA scaffold sequence. This work was supported by the US National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, EB010249 (S.R.J.).

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W.S., R.L.S. and S.R.J. conceived and designed the experiments, W.S. and R.L.S. performed experiments and analyzed data, and W.S., R.L.S. and S.R.J. wrote the manuscript.

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Correspondence to Samie R Jaffrey.

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S.R.J. and W.S. are authors of a patent application related to technology described in this manuscript.

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Song, W., Strack, R. & Jaffrey, S. Imaging bacterial protein expression using genetically encoded RNA sensors. Nat Methods 10, 873–875 (2013). https://doi.org/10.1038/nmeth.2568

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