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      Red-shifted click beetle luciferase mutant expands the multicolor bioluminescent palette for deep tissue imaging

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          Summary

          For in vivo multicolor bioluminescence applications, red and near-infrared signals are desirable over shorter wavelength signals because they are not as susceptible to light attenuation by blood and tissue. Herein, we describe the development of a new click beetle luciferase mutant, CBG2, with a red-shifted color emission. When paired with NH 2-NpLH2 luciferin, CBG2 (λ = 660 nm) and CBR2 (λ = 730 nm) luciferases can be used for simultaneous dual-color bioluminescence imaging in deep tissue. Using a spectral unmixing algorithm tool it is possible to distinguish each spectral contribution. Ultimately, this enzyme pair can expand the near-infrared bioluminescent toolbox to enable rapid visualization of multiple biological processes in deep tissue using a single substrate.

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          Highlights

          • •

            CBG2 is a new click beetle mutant derived from CBG99 luciferase

          • •

            It has a far red-shifted bioluminescent emission with NH 2-NpLH2 luciferin (λ = 660 nm)

          • •

            CBG2 with CBR2 luciferase and NH 2-NpLH2 allows fast acquisition of data in deep tissue

          Abstract

          Optical Imaging; Biological Services; Biophysics

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          Most cited references25

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          Bioimaging: second window for in vivo imaging.

          Andrew M. Smith, Michael C. Mancini, Shuming Nie (2010)
          Article 0 comments Cited 410 times – based on 0 reviews     Review now
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            Bioluminescence.

            Keith T. Wilson, Waylon J. Hastings (1997)
            Bioluminescence has evolved independently many times; thus the responsible genes are unrelated in bacteria, unicellular algae, coelenterates, beetles, fishes, and others. Chemically, all involve exergonic reactions of molecular oxygen with different substrates (luciferins) and enzymes (luciferases), resulting in photons of visible light (approximately 50 kcal). In addition to the structure of luciferan, several factors determine the color of the emissions, such as the amino acid sequence of the luciferase (as in beetles, for example) or the presence of accessory proteins, notably GFP, discovered in coelenterates and now used as a reporter of gene expression and a cellular marker. The mechanisms used to control the intensity and kinetics of luminescence, often emitted as flashes, also vary. Bioluminescence is credited with the discovery of how some bacteria, luminous or not, sense their density and regulate specific genes by chemical communication, as in the fascinating example of symbiosis between luminous bacteria and squid.
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              In Vivo Molecular Bioluminescence Imaging: New Tools and Applications.

              Laura Mezzanotte, Moniek van 't Root, Hacer Karatas … (2017)
              in vivo bioluminescence imaging (BLi) is an optical molecular imaging technique used to visualize molecular and cellular processes in health and diseases and to follow the fate of cells with high sensitivity using luciferase-based gene reporters. The high sensitivity of this technique arises from efficient photon production, followed by the reaction between luciferase enzymes and luciferin substrates. Novel discoveries and developments of luciferase reporters, substrates, and gene-editing techniques, and emerging fields of applications, promise a new era of deeper and more sensitive molecular imaging.
              Article 0 comments Cited 91 times – based on 0 reviews     Review now
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                Author and article information

                Contributors
                Laura Mezzanotte
                Journal
                Journal ID (nlm-ta): iScience
                Journal ID (iso-abbrev): iScience
                Title: iScience
                Publisher: Elsevier
                ISSN (Electronic): 2589-0042
                Publication date PMC-release: 26 December 2020
                Publication date Collection: 22 January 2021
                Publication date (Electronic): 26 December 2020
                Volume: 24
                Issue: 1
                Electronic Location Identifier: 101986
                Affiliations
                [1 ]Erasmus Medical Center, Radiology and Nuclear Medicine, Rotterdam, 3015 CE, the Netherlands
                [2 ]Erasmus Medical Center, Molecular Genetics, Rotterdam, 3015 CE, the Netherlands
                [3 ]Medres Medical Research GmBH, Cologne 50931, Germany
                [4 ]Promega Corporation, Madison, WI 53711, USA
                [5 ]Percuros B.V., Leiden, 1333 CL, the Netherlands
                [6 ]Chiesi Farmaceutici, Parma, 43122, Italy
                [7 ]Promega Biosciences Incorporated, San Luis Obispo, CA 93401, USA
                [8 ]CHUV Department of Oncology, University of Lausanne, 461011 Lausanne, Switzerland
                Author notes
                [∗ ]Corresponding author l.mezzanotte@ 123456erasmusmc.nl
                [9]

                Lead contact

                Article
                Publisher Item ID: S2589-0042(20)31183-4 Publisher ID: 101986
                DOI: 10.1016/j.isci.2020.101986
                PMC ID: 7811125
                PubMed ID: 33490896
                SO-VID: 780c4529-7d96-4c2c-8f2b-9b6375fc9cdb
                Copyright © © 2020 The Author(s)
                License:

                This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

                History
                Date received : 2 October 2020
                Date revision received : 7 December 2020
                Date accepted : 21 December 2020
                Categories
                Subject: Article

                Keywords: optical imaging,biological services,biophysics
                Data availability:
                Keywords: optical imaging, biological services, biophysics

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