A versatile bioluminescent probe with tunable color†

IF 4.2 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY RSC Chemical Biology Pub Date : 2024-09-20 DOI:10.1039/D4CB00101J
Zachary R. Torrey, Lila P. Halbers, Lorenzo Scipioni, Giulia Tedeschi, Michelle A. Digman and Jennifer A. Prescher
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Abstract

Bioluminescence is a powerful method for imaging in vivo, but applications at the microscale are far from routine. This is due, in part, to a lack of versatile tools for visualizing dynamic events. To address this void, we developed a new platform—Bioluminescence Resonance Energy mAKe over with a Fluorescence-Activating absorption-Shifting Tag (BREAKFAST). BREAKFAST features a bright luciferase combined with a chemogenetic tag (pFAST) for rapid color switching. In the presence of luciferin and a discrete fluorogenic ligand, signal is observed via resonance energy transfer. We evaluated spectral outputs with various fluorogens and established the utility of BREAKFAST for combined fluorescence and bioluminescence imaging. Dynamic, four-color visualization was achieved with sequential ligand addition and spectral phasor analysis. We further showed selective signal quenching with a dark fluorogen. Collectively, this work establishes a new method for bioluminescence imaging at the cellular scale and sets the stage for continued probe development.

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颜色可调的多功能生物发光探针。
生物发光是一种强大的体内成像方法,但在微观尺度上的应用还远未普及。部分原因是缺乏可视化动态事件的多功能工具。为了填补这一空白,我们开发了一种新的平台--带有荧光激活吸收位移标签(BREAKFAST)的生物发光共振能量 mAKe。BREAKFAST 的特点是将明亮的荧光素酶与化学标签(pFAST)相结合,以实现快速颜色转换。在荧光素和离散致荧光配体的作用下,通过共振能量转移观察信号。我们评估了各种荧光剂的光谱输出,并确定了 BREAKFAST 在荧光和生物发光联合成像中的实用性。通过依次添加配体和光谱相位分析,实现了动态四色可视化。我们还进一步展示了暗荧光剂的选择性信号淬灭。总之,这项工作为细胞尺度的生物发光成像建立了一种新方法,并为探针的继续开发奠定了基础。
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来源期刊
CiteScore
6.10
自引率
0.00%
发文量
128
审稿时长
10 weeks
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