Near-Infrared Fluorescent Proteins, Biosensors and Optogenetic Tools

近红外荧光蛋白、生物传感器和光遗传学工具

• 批准号: 10163867
• 负责人: Vladislav Verkhusha
• 金额: $ 44.12万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10163867
• 关键词: Animal Model; Animals; Biliverdine; Biochemistry; Biosensor; Cells; Chemicals; Complement; Detection; Development; Fluorescence; Fluorescence Resonance Energy Transfer; Gene Delivery; Heme; Hemoglobin; Image; Knowledge; Label; Light; Lighting; Mammalian Cell; Mammals; Melanins; Metabolic; Methods; Microscopy; Molecular; Molecular Weight; Nature; Optics; Organ; Pathology; Photochemistry; Physiology; Phytochrome; Process; Proteins; Reagent; Signal Transduction; Skin; Structure; Technology; Tissues; Visualization; Water; Whole Organism; base; chromophore; design; in vivo; in vivo imaging; non-invasive imaging; novel; optogenetics; photoactivation; tool

ABSTRACT Near-infrared light is favorable for imaging in mammalian tissues due to low absorbance of hemoglobin, melanin and water. Therefore, fluorescent proteins, biosensors and optogenetic constructs for optimal imaging, optical readout and light manipulation in mammals should have fluorescence and action spectra within the near-infrared window. Interestingly, natural bacterial phytochromes utilize the low molecular weight biliverdin, found in most mammalian tissues, as a photoreactive chromophore. Due to their near-infrared absorbance bacterial phytochromes are preferred templates for designing optical molecular tools for applications in mammals. Based on the analysis of the photochemistry and structure of bacterial phytochromes we suggest a variety of possible bacterial phytochrome-based fluorescent proteins, biosensors, and optogenetic tools. The design strategies and experimental considerations for such probes are proposed. Near-infrared fluorescent proteins and biosensors will extend the methods developed for conventional microscopy into a deep-tissue in vivo macroscopy including multicolor cell and tissue labeling, fluorescence resonance energy transfer, cell photoactivation and tracking, and detection of enzymatic activities and metabolites in tissues. The near-infrared optogenetic tools will allow noninvasive light-control of biochemistry and physiology of a living animal directly through the skin. Moreover, all bacterial phytochromes-based reagents will spectrally complement existing genetically encoded probes in the visible range. The planned probes also will expand our basic knowledge of photochemistry and light-induced signaling of phytochromes in nature. Availability of the near-infrared probes will further stimulate the development of novel in vivo imaging and light-manipulation technologies, optimization of strategies for gene delivery to specific cells and tissues in vivo, design of targeted noninvasive illumination, and refining optical readouts. Overall, this will result in a wide range of noninvasive studies of chemical and metabolic status, as well as molecular and cellular interactions in intact tissues and whole living mammals.

抽象的 由于血红蛋白吸光度低，近红外光有利于在哺乳动物组织中成像 黑色素和水。因此，用于最佳成像的荧光蛋白，生物传感器和光遗传构建体， 哺乳动物中的光学读数和光操纵应具有荧光和动作光谱 近红外窗户。有趣的是，天然细菌植物色素利用低分子量biliverdin， 在大多数哺乳动物组织中发现是一种光反应性发色团。由于它们的近红外吸光度 细菌植物色素是设计用于应用的光学分子工具的首选模板 哺乳动物。基于对细菌植物色素的光化学和结构的分析，我们建议 各种可能的细菌植物色素基荧光蛋白，生物传感器和光遗传学工具。这 提出了此类探针的设计策略和实验考虑。近红外荧光 蛋白质和生物传感器将将用于常规显微镜开发的方法扩展到深度组织中 体内宏观检查包括多色细胞和组织标记，荧光共振能量转移，细胞 光激活和跟踪，以及组织中的酶活性和代谢产物的检测。近红外 光遗传学工具将直接允许无创的生物化学和生理学的光照。 穿过皮肤。此外，所有基于细菌植物色素的试剂将在光谱中补充现有 可见范围内的遗传编码探针。计划的探针还将扩大我们对 植物色素自然界的光化学和光诱导的信号传导。近红外探针的可用性 将进一步刺激新型体内成像和光操作技术的发展，优化 基因输送到体内特定细胞和组织的策略，靶向非侵入性照明的设计， 并完善光学读数。总体而言，这将导致对化学和化学和 代谢状态，以及完整的组织和整个活哺乳动物中的分子和细胞相互作用。