Milestones in the development and implementation of FRET-based sensors of intracellular signals: A biological perspective of the history of FRET

Deal, Joshua; Pleshinger, D. J.; Johnson, Santina; Leavesley, Silas J.; Rich, Thomas C.

doi:10.1016/j.cellsig.2020.109769

Cited by 19 publications

(12 citation statements)

References 99 publications

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“…Genetically encoded fluorescent proteins (FPs)-based Förster resonance energy transfer (FRET) technology has been widely used for mapping temporal-spatial dynamics of intracellular biochemical events in living cells [1][2][3][4][5][6][7][8][9][10][11] "3-cube FRET" microscopy, referred to 3-cube FRET method is the most extensively applied approach for live-cell FRET quantification. [12][13][14][15] Three images are needed for E-FRET image: I DD (DA), image from donor channel with donor excitation; I AA (DA), image from acceptor channel with acceptor excitation; and I DA (DA), image from acceptor channel with donor excitation.…”

Section: Introductionmentioning

confidence: 99%

Measuring calibration factors by imaging a dish of cells expressing different tandem constructs plasmids

et al. 2021

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Three-cube Förster resonance energy transfer (FRET) method is the most extensively applied approach for live-cell FRET quantification. Reliable measurements of calibration factors are crucial for quantitative FRET measurement. We here proposed a modified TA-G method (termed as mTA-G) to simultaneously obtain the FRETsensitized quenching transition factor (G) and extinction coefficients ratio (γ) between donor and acceptor. mTA-G method includes four steps: ( 1) predetermining the ratio ranges of the sensitized emission of acceptor (F C ) to the donor excitation and donor channel image (I DD [(DA])) for all FRET plasmids; (2) culturing the cells which express every FRET plasmid in one dish respectively; (3) distinguishing and marking the cells expressing different FRET plasmids by detecting their F C /I DD (DA) values; (4) linearly fitting F C /I AA (DA) (acceptor excitation and acceptor channel image) to I DD (DA)/I AA (DA)for different kinds of cells. We implemented mTA-G method by imaging tandem constructs cells with different FRET efficiency cultured in one dish on different days, and obtained consistent G and γ values. mTA-G method not only circumvents switchover of different culture dishes but also keep the constant imaging conditions, exhibiting excellent robustness, and thus will expands the biological applications of quantitative FRET analysis in living cells.

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Section: Introductionmentioning

confidence: 99%

Measuring calibration factors by imaging a dish of cells expressing different tandem constructs plasmids

et al. 2021

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“…FRET (fluorescence resonance energy transfer) and FRAP (fluorescence recovery after photobleaching) both provide useful information about protein-lipid systems. FRET involves illuminating a fluorophore and measuring the resulting emission from another (donor and acceptor in Figure 3)so they have to be close enough to transfer energy, with the intensity following Förster's [13,14]. FRET is especially useful in estimating intramolecular distances and dynamic conformational changes perhaps as a function of property change measurements (e.g.…”

Section: Fluorescence Of Membrane-bound Peptidesmentioning

confidence: 99%

Spectroscopy of model-membrane liposome-protein systems: complementarity of linear dichroism, circular dichroism, fluorescence and SERS

Rodger

2021

Emerging Topics in Life Sciences

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A range of membrane models have been developed to study components of cellular systems. Lipid vesicles or liposomes are one such artificial membrane model which mimics many properties of the biological system: they are lipid bilayers composed of one or more lipids to which other molecules can associate. Liposomes are thus ideal to study the roles of cellular lipids and their interactions with other membrane components to understand a wide range of cellular processes including membrane disruption, membrane transport and catalytic activity. Although liposomes are much simpler than cellular membranes, they are still challenging to study and a variety of complementary techniques are needed. In this review article, we consider several currently used analytical methods for spectroscopic measurements of unilamellar liposomes and their interaction with proteins and peptides. Among the variety of spectroscopic techniques seeing increasing application, we have chosen to discuss: fluorescence based techniques such as FRET (fluorescence resonance energy transfer) and FRAP (fluorescence recovery after photobleaching), that are used to identify localisation and dynamics of molecules in the membrane; circular dichroism (CD) and linear dichroism (LD) for conformational and orientation changes of proteins on membrane binding; and SERS (Surface Enhanced Raman Spectroscopy) as a rapidly developing ultrasensitive technique for site-selective molecular characterisation. The review contains brief theoretical basics of the listed techniques and recent examples of their successful applications for membrane studies.

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“…Förster resonance energy transfer (FRET) is the radiationless transfer of excitation energy occurring between the singlet excited state of the donor (D) and the ground state of the acceptor (A) through dipole-dipole interaction [ 13 ]. This process plays a very important role in prolific areas of science and technology, including the study of protein-protein interactions, the intramolecular distance between selected sites of biospecies, the study and design of optical properties of functional materials, nanostructures, molecular mono- and multilayers, interfaces, aggregates, plasmonically enhanced broadband emission or some ordered systems [ 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 ].…”

Section: Introductionmentioning

confidence: 99%

New Core-Shell Nanostructures for FRET Studies: Synthesis, Characterization, and Quantitative Analysis

Synak

Adamska

Kułak

et al. 2022

IJMS

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This work describes the synthesis and characterization of new core-shell material designed for Förster resonance energy transfer (FRET) studies. Synthesis, structural and optical properties of core-shell nanostructures with a large number of two kinds of fluorophores bound to the shell are presented. As fluorophores, strongly fluorescent rhodamine 101 and rhodamine 110 chloride were selected. The dyes exhibit significant spectral overlap between acceptor absorption and donor emission spectra, which enables effective FRET. Core-shell nanoparticles strongly differing in the ratio of donors to acceptor numbers were prepared. This leads to two different interesting cases: typical single-step FRET or multistep energy migration preceding FRET. The single-step FRET model that was designed and presented by some of us recently for core-shell nanoparticles is herein experimentally verified. Very good agreement between the analytical expression for donor fluorescence intensity decay and experimental data was obtained, which confirmed the correctness of the model. Multistep energy migration between donors preceding the final transfer to the acceptor can also be successfully described. In this case, however, experimental data are compared with the results of Monte Carlo simulations, as there is no respective analytical expression. Excellent agreement in this more general case evidences the usefulness of this numerical method in the design and prediction of the properties of the synthesized core-shell nanoparticles labelled with multiple and chemically different fluorophores.

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Milestones in the development and implementation of FRET-based sensors of intracellular signals: A biological perspective of the history of FRET

Cited by 19 publications

References 99 publications

Measuring calibration factors by imaging a dish of cells expressing different tandem constructs plasmids

Measuring calibration factors by imaging a dish of cells expressing different tandem constructs plasmids

Spectroscopy of model-membrane liposome-protein systems: complementarity of linear dichroism, circular dichroism, fluorescence and SERS

New Core-Shell Nanostructures for FRET Studies: Synthesis, Characterization, and Quantitative Analysis

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