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

Giorgia Zambito; MP Hall; MG Wood; Natasa Gaspar; Yanto Ridwan; FF Stellari; C Shi; TA Kirkland; LP Encell; Clemens Löwik; Laura Mezzanotte

doi:10.1016/j.isci.2020.101986

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

Giorgia Zambito, MP Hall, MG Wood, Natasa Gaspar, Yanto Ridwan, FF Stellari, C Shi, TA Kirkland, LP Encell, Clemens Löwik, Laura Mezzanotte

Research output: Contribution to journal › Article › Academic › peer-review

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Abstract

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.

Original language	English
Article number	101986
Journal	iScience
Volume	24
Issue number	1
DOIs	https://doi.org/10.1016/j.isci.2020.101986
Publication status	Published - 22 Jan 2021

Bibliographical note

Funding Information:
We acknowledge the funding for this work provided by the European Commission under the H2020-MSCA-RISE award grant number 777682 ( CANCER ) and under the H2020-MSCA-ITN award, grant number 675743 (ISPIC). This work was supported by the Applied Molecular Imaging Erasmus MC (AMIE) facility.

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© 2020 The Author(s)

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

abstract = "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. ",

author = "Giorgia Zambito and MP Hall and MG Wood and Natasa Gaspar and Yanto Ridwan and FF Stellari and C Shi and TA Kirkland and LP Encell and Clemens L{\"o}wik and Laura Mezzanotte",

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AU - Zambito, Giorgia

AU - Hall, MP

AU - Wood, MG

AU - Gaspar, Natasa

AU - Ridwan, Yanto

AU - Stellari, FF

AU - Shi, C

AU - Kirkland, TA

AU - Encell, LP

AU - Löwik, Clemens

AU - Mezzanotte, Laura

N1 - Funding Information: We acknowledge the funding for this work provided by the European Commission under the H2020-MSCA-RISE award grant number 777682 ( CANCER ) and under the H2020-MSCA-ITN award, grant number 675743 (ISPIC). This work was supported by the Applied Molecular Imaging Erasmus MC (AMIE) facility. Publisher Copyright: © 2020 The Author(s)

PY - 2021/1/22

Y1 - 2021/1/22

N2 - 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.

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

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