2. Academic Validation
  3. Five colour variants of bright luminescent protein for real-time multicolour bioimaging

Five colour variants of bright luminescent protein for real-time multicolour bioimaging

  • Nat Commun. 2016 Dec 14:7:13718. doi: 10.1038/ncomms13718.
Kazushi Suzuki 1,  Taichi Kimura 2,  Hajime Shinoda 1,  Guirong Bai 3,  Matthew J Daniels 4,  Yoshiyuki Arai 1 2 3,  Masahiro Nakano 1 2 3,  Takeharu Nagai 1 2 3
Affiliations

Affiliations

  • 1 Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan.
  • 2 Department of Biotechnology, School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan.
  • 3 Department of Biomolecular Science and Engineering, The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki 567-0047, Japan.
  • 4 BHF Centre for Regenerative Medicine, Division of Cardiovascular Medicine, West Wing Level 6, John Radcliffe Hospital, Oxford OX3 9DU, UK.
PMID: 27966527 DOI: 10.1038/ncomms13718
Abstract

Luminescence imaging has gained attention as a promising bio-imaging modality in situations where fluorescence imaging cannot be applied. However, wider application to multicolour and dynamic imaging is limited by the lack of bright luminescent proteins with emissions across the visible spectrum. Here we report five new spectral variants of the bright luminescent protein, enhanced Nano-lantern (eNL), made by concatenation of the brightest luciferase, NanoLuc, with various colour hues of fluorescent proteins. eNLs allow five-colour live-cell imaging, as well as detection of single protein complexes and even single molecules. We also develop an eNL-based CA2+ indicator with a 500% signal change, which can image spontaneous CA2+ dynamics in cardiomyocyte and neural cell models. These eNL probes facilitate not only multicolour imaging in living cells but also sensitive imaging of a wide repertoire of proteins, even at very low expression levels.

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