title: Five colour variants of bright luminescent protein for real-time multicolour bioimaging.
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.
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan.
authors.forename: Kazushi
authors.initials: K
authors.lastname: Suzuki
authors.affiliation: Department of Biotechnology, School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan.
authors.forename: Taichi
authors.initials: T
authors.lastname: Kimura
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan.
authors.forename: Hajime
authors.initials: H
authors.lastname: Shinoda
authors.affiliation: Department of Biomolecular Science and Engineering, The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki 567-0047, Japan.
authors.forename: Guirong
authors.initials: G
authors.lastname: Bai
authors.affiliation: BHF Centre for Regenerative Medicine, Division of Cardiovascular Medicine, West Wing Level 6, John Radcliffe Hospital, Oxford OX3 9DU, UK.
authors.forename: Matthew J
authors.identifier: http://orcid.org/0000-0002-9250-6214
authors.initials: MJ
authors.lastname: Daniels
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan.|Department of Biotechnology, School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan.|Department of Biomolecular Science and Engineering, The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki 567-0047, Japan.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Arai
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan.|Department of Biotechnology, School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan.|Department of Biomolecular Science and Engineering, The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki 567-0047, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Nakano
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan.|Department of Biotechnology, School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan.|Department of Biomolecular Science and Engineering, The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki 567-0047, Japan.
authors.forename: Takeharu
authors.initials: T
authors.lastname: Nagai
mesh_terms: D002118:Calcium / Q000032:analysis; D020013:Calcium Signaling; D003116:Color; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis / Q000737:chemistry*; D032383:Myocytes, Cardiac / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D061848:Optical Imaging / Q000379:methods
chemical_list: D008164:Luminescent Proteins; D002118:Calcium