Zambito Giorgia, Hall Mary P, Wood Monika G, Gaspar Natasa, Ridwan Yanto, Stellari Fabio F, Shi Ce, Kirkland Thomas A, Encell Lance P, Löwik Clemens, Mezzanotte Laura
Erasmus Medical Center, Radiology and Nuclear Medicine, Rotterdam, 3015 CE, the Netherlands.
Erasmus Medical Center, Molecular Genetics, Rotterdam, 3015 CE, the Netherlands.
iScience. 2020 Dec 26;24(1):101986. doi: 10.1016/j.isci.2020.101986. eCollection 2021 Jan 22.
For 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-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.
对于多色生物发光应用而言，相比于较短波长的信号，红色和近红外信号更受青睐，因为它们不太容易受到血液和组织的光衰减影响。在此，我们描述了一种新的叩头虫荧光素酶突变体CBG2的开发，其具有红移的颜色发射。当与NH-NpLH2荧光素配对时，CBG2（λ = 660 nm）和CBR2（λ = 730 nm）荧光素酶可用于深层组织中的同步双色生物发光成像。使用光谱解混算法工具可以区分每个光谱贡献。最终，这对酶可以扩展近红外生物发光工具箱，以便使用单一底物快速可视化深层组织中的多个生物学过程。
