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Dual Reporter Bioluminescence Imaging with NanoLuc and Firefly Luciferase

  • Anne E. Gibbons
  • Kathryn E. Luker
  • Gary D. LukerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1790)

Abstract

Bioluminescence imaging is a powerful, broadly utilized method for noninvasive imaging studies in cell-based assays and small animal models of normal physiology and multiple diseases. In combination with molecular engineering of cells and entire organisms using luciferase enzymes, bioluminescence imaging has enabled novel applications including studies of protein-protein interactions, ligand-receptor interactions, cell trafficking, and drug targeting in mouse models. We describe use of a novel luciferase enzyme derived from Oplophorus gracilirostris, NanoLuc, in cell-based assays bioluminescence imaging of tumor-bearing mice. We also combine NanoLuc with another luciferase enzyme, firefly luciferase, to image multiple signal transduction events in one imaging session.

Key words

NanoLuc Bioluminescence imaging Firefly luciferase Oplophorus gracilirostris 

References

  1. 1.
    Luker K, Luker G (2010) Bioluminescence imaging of reporter mice for studies of infection and inflammation. Antivir Res 86(1):93–100CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Luker K, Smith M, Luker G et al (2004) Kinetics of regulated protein-protein interactions revealed with firefly luciferase complementation imaging in cells and living animals. Proc Natl Acad Sci U S A 101(33):12288–12293CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Zhang L, Lee K, Bhojani M et al (2007) Molecular imaging of Akt kinase activity. Nat Med 13(9):1114–1119CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Wang H, Cao F, De A et al (2009) Trafficking mesenchymal stem cell engraftment and differentiation in tumor-bearing mice by bioluminescence imaging. Stem Cells 27(7):1548–1558CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    de Almeida P, van Rappard J, Wu J (2011) In vivo bioluminescence for tracking cell fate and function. Am J Physiol Heart Circ Physiol 301(3):H663–H671CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Luker K, Mihalko L, Schmidt B, Lewin S et al (2012) In vivo imaging of ligand receptor binding with Gaussia luciferase complementation. Nat Med 18(1):172–177CrossRefGoogle Scholar
  7. 7.
    Zinn K, Chaudhuri T, Szafran A et al (2008) Noninvasive bioluminescence imaging in small animals. ILAR J 49(1):103–115CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Bhaumik S, Gambhir S (2002) Optical imaging of Renilla luciferase reporter gene expression in living mice. Proc Natl Acad Sci U S A 99(1):377–382CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Tannous B, Kim D, Fernandez J et al (2005) Codon-optimized Gaussia luciferase cDNA for mammalian gene expression in culture and in vivo. Mol Ther 11(3):435–443CrossRefPubMedGoogle Scholar
  10. 10.
    Venisnik K, Olafsen T, Gambhir S (2007) Fusion of Gaussia luciferase to an engineered anti-carcinoembryonic antigen (CEA) antibody for in vivo optical imaging. Mol Imaging Biol 9(5):267–277CrossRefPubMedGoogle Scholar
  11. 11.
    Hall M, Unch J, Binkowski B et al (2012) Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chem Biol 7(11):1848–1857CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Stacer A, Nyati S, Moudgil P et al (2013) NanoLuc reporter for dual luciferase imaging in living animals. Mol Imaging 12(7):1–13CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Nyati S, Schinske K, Ray D et al (2011) Molecular imaging of TGFβ-induced Smad2/3 phosphorylation reveals a role for receptor tyrosine kinases in modulating TGFβ signaling. Clin Cancer Res 17(23):7424–7439CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Anne E. Gibbons
    • 1
  • Kathryn E. Luker
    • 1
  • Gary D. Luker
    • 1
    • 2
    • 3
    Email author
  1. 1.Department of Radiology, Center for Molecular ImagingUniversity of MichiganAnn ArborUSA
  2. 2.Department of Biomedical EngineeringUniversity of MichiganAnn ArborUSA
  3. 3.Department of Microbiology and ImmunologyUniversity of MichiganAnn ArborUSA

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