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Red-shifted luciferase–luciferin pairs for enhanced bioluminescence imaging

Nature Methods volume 14, pages 971974 (2017) | Download Citation

Abstract

Red-shifted bioluminescence reporters are desirable for biological imaging. We describe the development of red-shifted luciferins based on synthetic coelenterazine analogs and corresponding mutants of NanoLuc that enable bright bioluminescence. One pair in particular showed superior in vitro and in vivo sensitivity over commonly used bioluminescence reporters. We adapted this pair to develop a bioluminescence resonance-energy-based Antares reporter called Antares2, which offers improved signal from deep tissues.

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Acknowledgements

pNCS-Antares and pcDNA3-ReNL were gifts from M. Lin and T. Nagai (Addgene plasmids 74279 and 85203), respectively. We thank the University of California, Riverside, the National Institutes of Health (R01GM118675 and R21EB021651), and the National Science Foundation (CHE-1351933) for financial support. We thank T.M. Truong for reading and revising the manuscript.

Author information

Affiliations

  1. Department of Chemistry, University of California Riverside, Riverside, California, USA.

    • Hsien-Wei Yeh
    • , Ao Ji
    •  & Hui-wang Ai
  2. Department of Molecular Physiology and Biological Physics and Center for Membrane and Cell Physiology, University of Virginia School of Medicine, Charlottesville, Virginia, USA.

    • Hsien-Wei Yeh
    •  & Hui-wang Ai
  3. Department of Cell Biology and Neuroscience, University of California Riverside, Riverside, California, USA.

    • Omran Karmach
    •  & Manuela M Martins-Green
  4. Institute for Integrative Genome Biology, University of California Riverside, Riverside, California, USA.

    • David Carter
    • , Manuela M Martins-Green
    •  & Hui-wang Ai

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Contributions

H.-w.A. conceived and supervised the entire project. H.-w.A. and M.M.M.-G. supervised the mouse imaging experiments. H.-W.Y. performed synthetic, protein engineering, and in vitro and in cellulo characterization experiments. H.-W.Y. and O.K. performed the mouse imaging experiments. A.J. assisted H.-W.Y. in completing compound synthesis. D.C. assisted in the mouse imaging experiments. H.-W.Y. and H.-w.A. analyzed the data and wrote the manuscript.

Competing interests

UC Riverside has filed a provisional patent application in the USA (No. 62/382,255) that is partially based on results described in the manuscript. H.-w.A. and H.-W.Y. are listed as coinventors.

Corresponding author

Correspondence to Hui-wang Ai.

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DOI

https://doi.org/10.1038/nmeth.4400

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