Red-shifted click beetle luciferase mutant expands the multicolor bioluminescent palette for deep tissue imaging
Giorgia Zambito,
Mary P. Hall,
Monika G. Wood,
Natasa Gaspar,
Yanto Ridwan,
Fabio F. Stellari,
Ce Shi,
Thomas A. Kirkland,
Lance P. Encell,
Clemens Löwik,
Laura Mezzanotte
Affiliations
Giorgia Zambito
Erasmus Medical Center, Radiology and Nuclear Medicine, Rotterdam, 3015 CE, the Netherlands; Erasmus Medical Center, Molecular Genetics, Rotterdam, 3015 CE, the Netherlands; Medres Medical Research GmBH, Cologne 50931, Germany
Mary P. Hall
Promega Corporation, Madison, WI 53711, USA
Monika G. Wood
Promega Corporation, Madison, WI 53711, USA
Natasa Gaspar
Erasmus Medical Center, Radiology and Nuclear Medicine, Rotterdam, 3015 CE, the Netherlands; Erasmus Medical Center, Molecular Genetics, Rotterdam, 3015 CE, the Netherlands; Percuros B.V., Leiden, 1333 CL, the Netherlands
Yanto Ridwan
Erasmus Medical Center, Radiology and Nuclear Medicine, Rotterdam, 3015 CE, the Netherlands; Erasmus Medical Center, Molecular Genetics, Rotterdam, 3015 CE, the Netherlands
Fabio F. Stellari
Chiesi Farmaceutici, Parma, 43122, Italy
Ce Shi
Promega Biosciences Incorporated, San Luis Obispo, CA 93401, USA
Thomas A. Kirkland
Promega Biosciences Incorporated, San Luis Obispo, CA 93401, USA
Lance P. Encell
Promega Corporation, Madison, WI 53711, USA
Clemens Löwik
Erasmus Medical Center, Radiology and Nuclear Medicine, Rotterdam, 3015 CE, the Netherlands; Erasmus Medical Center, Molecular Genetics, Rotterdam, 3015 CE, the Netherlands; CHUV Department of Oncology, University of Lausanne, 461011 Lausanne, Switzerland
Laura Mezzanotte
Erasmus Medical Center, Radiology and Nuclear Medicine, Rotterdam, 3015 CE, the Netherlands; Erasmus Medical Center, Molecular Genetics, Rotterdam, 3015 CE, the Netherlands; Corresponding author
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 NH2-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.
