探索基于 FRET 的分子传感器:新兴应用领域的设计策略和最新进展。

IF 12.1 1区 工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biotechnology advances Pub Date : 2024-10-16 DOI:10.1016/j.biotechadv.2024.108466
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引用次数: 0

摘要

探测生物体内的生物过程是一项艰巨的任务,需要专门的分析设备才能对重要生理参数的实时变化提供独到的见解。经典的生物化学方法极大地帮助了我们对重要生物过程调控机制的理解。然而,这些方法通常不足以研究瞬时分子事件,因为它们主要关注的是最终结果。荧光共振能量转移(FRET)显微镜是一种有效的工具,可利用精心构建的传感器,非侵入式地探索蛋白质之间的实时动态相互作用和各种生化信号事件。由于其多功能性,基于 FRET 的传感器能够对大量生物变量进行快速、标准化的评估,促进了高通量研究和具有卓越时空分辨率的精确亚细胞测量。这篇综述首先简要介绍了 FRET 理论,讨论了在化学生物学研究的不同传感模式中可用作标记的荧光分子,然后概述了目前用于量化 FRET 的成像技术,重点介绍了这些技术的优势和不足。文章还讨论了可用于构建 FRET 支架的各种供体-受体组合。具体来说,这篇综述深入介绍了基于 FRET 的传感器的最新实时生物成像应用,并讨论了此类设备的常见架构。此外,还讨论了具有复用功能的 FRET 系统和用于双/多分析物检测的多步骤 FRET 协议。此外,还提到了这一令人兴奋的领域未来的研究方向,以及面临的障碍和机遇。
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Exploring the landscape of FRET-based molecular sensors: Design strategies and recent advances in emerging applications
Probing biological processes in living organisms that could provide one-of-a-kind insights into real-time alterations of significant physiological parameters is a formidable task that calls for specialized analytic devices. Classical biochemical methods have significantly aided our understanding of the mechanisms that regulate essential biological processes. These methods, however, are typically insufficient for investigating transient molecular events since they focus primarily on the end outcome. Fluorescence resonance energy transfer (FRET) microscopy is a potent tool used for exploring non-invasively real-time dynamic interactions between proteins and a variety of biochemical signaling events using sensors that have been meticulously constructed. Due to their versatility, FRET-based sensors have enabled the rapid and standardized assessment of a large array of biological variables, facilitating both high-throughput research and precise subcellular measurements with exceptional temporal and spatial resolution. This review commences with a brief introduction to FRET theory and a discussion of the fluorescent molecules that can serve as tags in different sensing modalities for studies in chemical biology, followed by an outlining of the imaging techniques currently utilized to quantify FRET highlighting their strengths and shortcomings. The article also discusses the various donor-acceptor combinations that can be utilized to construct FRET scaffolds. Specifically, the review provides insights into the latest real-time bioimaging applications of FRET-based sensors and discusses the common architectures of such devices. There has also been discussion of FRET systems with multiplexing capabilities and multi-step FRET protocols for use in dual/multi-analyte detections. Future research directions in this exciting field are also mentioned, along with the obstacles and opportunities that lie ahead.
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来源期刊
Biotechnology advances
Biotechnology advances 工程技术-生物工程与应用微生物
CiteScore
25.50
自引率
2.50%
发文量
167
审稿时长
37 days
期刊介绍: Biotechnology Advances is a comprehensive review journal that covers all aspects of the multidisciplinary field of biotechnology. The journal focuses on biotechnology principles and their applications in various industries, agriculture, medicine, environmental concerns, and regulatory issues. It publishes authoritative articles that highlight current developments and future trends in the field of biotechnology. The journal invites submissions of manuscripts that are relevant and appropriate. It targets a wide audience, including scientists, engineers, students, instructors, researchers, practitioners, managers, governments, and other stakeholders in the field. Additionally, special issues are published based on selected presentations from recent relevant conferences in collaboration with the organizations hosting those conferences.
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