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 由于血红蛋白的吸光度较低，近红外光有利于哺乳动物组织的成像， 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学结构， 哺乳动物的光学读出和光操纵应该在范围内具有荧光和作用光谱 近红外窗口。有趣的是，天然细菌光敏色素利用低分子量胆绿素， 作为光反应性发色团存在于大多数哺乳动物组织中。由于它们的近红外吸收 细菌光敏色素是设计用于以下应用的光学分子工具的首选模板 哺乳动物。基于对细菌光敏色素的光化学和结构的分析，我们建议 各种可能的基于细菌光敏色素的荧光蛋白、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因素。近红外荧光 蛋白质和生物传感器将把传统显微镜开发的方法扩展到深层组织 活体肉眼观察，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，以及组织中酶活性和代谢物的检测。近红外线 光遗传学工具将允许直接对活体动物的生物化学和生理学进行无创光控制 通过皮肤。此外，所有基于细菌光敏色素的试剂都将在光谱上补充现有的 可见光范围内的基因编码探针。计划中的探索还将扩展我们的基本知识 自然界光敏色素的光化学和光诱导信号传导。近红外探头的可用性 将进一步刺激新型体内成像和光操纵技术、优化的发展 将基因传递到体内特定细胞和组织的策略，有针对性的非侵入性照明的设计， 并改进光学读数。总体而言，这将导致广泛的化学和非侵入性研究 代谢状态，以及完整组织和整个活哺乳动物中的分子和细胞相互作用。