Abstract
A common application of fluorescent proteins is to label whole cells, but many RFPs are cytotoxic when used with standard high-level expression systems. We engineered a rapidly maturing tetrameric fluorescent protein called DsRed-Express2 that has minimal cytotoxicity. DsRed-Express2 exhibits strong and stable expression in bacterial and mammalian cells, and it outperforms other available RFPs with regard to photostability and phototoxicity.
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Acknowledgements
We thank for technical assistance J. Jureller and members of the Institute for Biophysical Dynamics NanoBiology Facility at The University of Chicago, and members of The University of Chicago Flow Cytometry Facility and The Indiana University Simon Cancer Center Flow Cytometry Resource Facility. We also thank R. Tsien (University of California at San Diego) for providing mCherry and tdTomato constructs; J. Wiedenmann (University of Ulm) for providing RFP611 construct; S. Crosson for use of the spectrophotometer; G. Elder for helpful discussion; and members of the Glick and Keenan labs for feedback on the manuscript. This work was supported by US National Institutes of Health grants R01 EB008087 (to B.S.G. and R.J.K.), T32 GM007183 (to R.L.S.), and R01 HL56416, R01 HL67384 and a project in P01 HL53586 (to H.E.B.).
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R.L.S., D.E.S., D.B., R.J.K. and B.S.G. are authors on a license agreement with Clontech to distribute DsRed-Express2.
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Strack, R., Strongin, D., Bhattacharyya, D. et al. A noncytotoxic DsRed variant for whole-cell labeling. Nat Methods 5, 955–957 (2008). https://doi.org/10.1038/nmeth.1264
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DOI: https://doi.org/10.1038/nmeth.1264
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