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A noncytotoxic DsRed variant for whole-cell labeling

Nature Methods volume 5, pages 955957 (2008) | Download Citation

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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Protein Data Bank

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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.).

Author information

Author notes

    • Rita L Strack
    •  & Daniel E Strongin

    These authors contributed equally to this work.

Affiliations

  1. Department of Biochemistry and Molecular Biology, The University of Chicago, Gordon Center W238, Chicago, Illinois 60637, USA.

    • Rita L Strack
    •  & Robert J Keenan
  2. Department of Molecular Genetics and Cell Biology, and Institute for Biophysical Dynamics, The University of Chicago, 920 East 58th Street, Chicago, Illinois 60637, USA.

    • Daniel E Strongin
    • , Dibyendu Bhattacharyya
    • , Allison Berman
    •  & Benjamin S Glick
  3. Walther Oncology Center, Indiana University School of Medicine, 950 West Walnut Street, Room 302, Indianapolis, Indiana 46202, USA.

    • Wen Tao
    •  & Hal E Broxmeyer

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Competing interests

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.

Corresponding author

Correspondence to Benjamin S Glick.

Supplementary information

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    Supplementary Text and Figures

    Supplementary Figures 1–8, Supplementary Tables 1–5, Supplementary Results, Supplementary Methods

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DOI

https://doi.org/10.1038/nmeth.1264

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