FRET as a biomolecular research tool-understanding its potential while avoiding pitfalls
FRET as a biomolecular research tool-understanding its potential while avoiding pitfalls
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DOI：
10.1038/s41592-019-0530-8
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发表时间：
2019-09-01
期刊：
影响因子：
48
通讯作者：
Medintz, Igor L.
中科院分区：
文献类型：
--
作者：
Algar, W. Russ;Hildebrandt, Niko;Medintz, Igor L.
福斯特共振能量转移（FRET）技术的应用日益广泛。然而，不同的FRET技术并没有一致地应用，也没有统一的结果，这使得实施和再现FRET实验具有挑战性。我们讨论了设计和评估合奏FRET实验的重要考虑因素。除了FRET基础上的引物，我们提供了实验设计选择的指导方针，如供体-受体对，仪器和标记化学;选择对照实验，明确证明FRET和验证实验提供有关生物分子过程的有意义的数据;分析原始数据和评估结果;并以易于再现的方式报告数据和实验细节。一些考虑也给出了FRET测定和FRET成像，特别是与荧光蛋白。我们的目标是激励和授权所有生物学家考虑FRET的强大的研究工具，它可以。
The applications of Forster resonance energy transfer (FRET) grow with each year. However, different FRET techniques are not applied consistently, nor are results uniformly presented, which makes implementing and reproducing FRET experiments challenging. We discuss important considerations for designing and evaluating ensemble FRET experiments. Alongside a primer on FRET basics, we provide guidelines for making experimental design choices such as the donor-acceptor pair, instrumentation and labeling chemistries; selecting control experiments to unambiguously demonstrate FRET and validate that the experiments provide meaningful data about the biomolecular process in question; analyzing raw data and assessing the results; and reporting data and experimental details in a manner that easily allows for reproducibility. Some considerations are also given for FRET assays and FRET imaging, especially with fluorescent proteins. Our goal is to motivate and empower all biologists to consider FRET for the powerful research tool it can be.