Toolkit/miRFP670-miRFP720 FRET pair

miRFP670-miRFP720 FRET pair

Multi-Component Switch·Research·Since 2018

Also known as: fully NIR FRET pair miRFP670-miRFP720

Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.

Summary

The miRFP670-miRFP720 FRET pair is a fully near-infrared genetically encoded Förster resonance energy transfer pair used to construct biosensors. It enables multiplexed biosensor imaging that is compatible with CFP-YFP imaging channels and blue-green optogenetic tools.

Usefulness & Problems

Why this is useful

This FRET pair is useful for building biosensors that can be imaged in the near-infrared while leaving visible-spectrum channels available for other reporters or actuators. In the cited study, it enabled simultaneous observation of Rac1 activity during optogenetic manipulation of Rac1 and supported multiplexed imaging of Rho GTPase signaling.

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This is a fully near-infrared FRET pair used to build biosensors. Its main role in the abstract is to enable multiplexed imaging without conflicting with visible-spectrum imaging and optogenetic channels.

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designing fully near-infrared FRET biosensors

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multiplex imaging with CFP-YFP biosensors and blue-green optogenetic tools

Problem solved

It addresses spectral crowding that can hinder direct multiplex imaging and simultaneous optogenetic control when conventional visible-spectrum reporters are used. The evidence specifically supports compatibility with CFP-YFP imaging and blue-green optogenetic tools.

Source:

It addresses spectral crowding that otherwise makes direct multiplex imaging and simultaneous optogenetic control difficult.

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spectral overlap between biosensors and optogenetic tools

Published Workflows

Direct multiplex imaging and optogenetics of Rho GTPases enabled by near-infrared FRET

2018

Objective: Develop a near-infrared FRET-based Rac1 biosensor and use it together with visible-spectrum biosensors and optogenetic control to directly image and perturb Rho GTPase signaling without problematic spectral overlap.

Why it works: The abstract states that the red-shifted miRFP720 and the fully NIR miRFP670-miRFP720 FRET pair enabled biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools, allowing simultaneous readout and perturbation.

Rac1 GTPase activity sensingRhoA-Rac1 antagonismRac1-GDI binding coordinationupstream Rac1 activationnear-infrared FRET biosensor designmultiplexed fluorescence imagingoptogenetic perturbation

Taxonomy & Function

Primary hierarchy

Mechanism Branch

Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.

Target processes

No target processes tagged yet.

Input: Light

Implementation Constraints

Implementation requires a genetically encoded FRET biosensor architecture containing both miRFP670 and miRFP720. Practical use also requires fluorescence imaging capable of detecting near-infrared FRET signals; the evidence does not provide additional construct design, cofactor, or expression-system details.

The supplied evidence describes the pair as a component for biosensor construction rather than a standalone pathway-specific sensor, so biological specificity depends on the surrounding biosensor design. The provided literature excerpt does not report quantitative photophysical parameters, dynamic range, maturation behavior, or validation beyond the cited multiplexed Rho GTPase applications.

Validation

Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication

Supporting Sources

Ranked Claims

Claim 1application demosupports2018Source 1needs review

The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.

and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
Claim 2application demosupports2018Source 1needs review

The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.

and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
Claim 3application demosupports2018Source 1needs review

The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.

and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
Claim 4application demosupports2018Source 1needs review

The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.

and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
Claim 5application demosupports2018Source 1needs review

The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.

and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
Claim 6application demosupports2018Source 1needs review

The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.

and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
Claim 7application demosupports2018Source 1needs review

The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.

and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
Claim 8application demosupports2018Source 1needs review

The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.

and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
Claim 9application demosupports2018Source 1needs review

The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.

and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
Claim 10biological observationsupports2018Source 1needs review

Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules.

showed that Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules
Claim 11biological observationsupports2018Source 1needs review

Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules.

showed that Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules
Claim 12biological observationsupports2018Source 1needs review

Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules.

showed that Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules
Claim 13biological observationsupports2018Source 1needs review

Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules.

showed that Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules
Claim 14biological observationsupports2018Source 1needs review

Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules.

showed that Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules
Claim 15biological observationsupports2018Source 1needs review

Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules.

showed that Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules
Claim 16biological observationsupports2018Source 1needs review

Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules.

showed that Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules
Claim 17biological observationsupports2018Source 1needs review

Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules.

showed that Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules
Claim 18biological observationsupports2018Source 1needs review

Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules.

showed that Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules
Claim 19biological observationsupports2018Source 1needs review

Using the multiplexed imaging setup, the authors directly observed and quantified antagonism between RhoA and Rac1 that depended on the RhoA-downstream effector ROCK.

We directly observed and quantified antagonism between RhoA and Rac1 dependent on the RhoA-downstream effector ROCK
Claim 20biological observationsupports2018Source 1needs review

Using the multiplexed imaging setup, the authors directly observed and quantified antagonism between RhoA and Rac1 that depended on the RhoA-downstream effector ROCK.

We directly observed and quantified antagonism between RhoA and Rac1 dependent on the RhoA-downstream effector ROCK
Claim 21biological observationsupports2018Source 1needs review

Using the multiplexed imaging setup, the authors directly observed and quantified antagonism between RhoA and Rac1 that depended on the RhoA-downstream effector ROCK.

We directly observed and quantified antagonism between RhoA and Rac1 dependent on the RhoA-downstream effector ROCK
Claim 22biological observationsupports2018Source 1needs review

Using the multiplexed imaging setup, the authors directly observed and quantified antagonism between RhoA and Rac1 that depended on the RhoA-downstream effector ROCK.

We directly observed and quantified antagonism between RhoA and Rac1 dependent on the RhoA-downstream effector ROCK
Claim 23biological observationsupports2018Source 1needs review

Using the multiplexed imaging setup, the authors directly observed and quantified antagonism between RhoA and Rac1 that depended on the RhoA-downstream effector ROCK.

We directly observed and quantified antagonism between RhoA and Rac1 dependent on the RhoA-downstream effector ROCK
Claim 24biological observationsupports2018Source 1needs review

Using the multiplexed imaging setup, the authors directly observed and quantified antagonism between RhoA and Rac1 that depended on the RhoA-downstream effector ROCK.

We directly observed and quantified antagonism between RhoA and Rac1 dependent on the RhoA-downstream effector ROCK
Claim 25biological observationsupports2018Source 1needs review

Using the multiplexed imaging setup, the authors directly observed and quantified antagonism between RhoA and Rac1 that depended on the RhoA-downstream effector ROCK.

We directly observed and quantified antagonism between RhoA and Rac1 dependent on the RhoA-downstream effector ROCK
Claim 26biological observationsupports2018Source 1needs review

Using the multiplexed imaging setup, the authors directly observed and quantified antagonism between RhoA and Rac1 that depended on the RhoA-downstream effector ROCK.

We directly observed and quantified antagonism between RhoA and Rac1 dependent on the RhoA-downstream effector ROCK
Claim 27biological observationsupports2018Source 1needs review

Using the multiplexed imaging setup, the authors directly observed and quantified antagonism between RhoA and Rac1 that depended on the RhoA-downstream effector ROCK.

We directly observed and quantified antagonism between RhoA and Rac1 dependent on the RhoA-downstream effector ROCK
Claim 28tool capabilitysupports2018Source 1needs review

miRFP720 and the miRFP670-miRFP720 fully near-infrared FRET pair enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.

Here we report the most red-shifted monomeric near-infrared (NIR) fluorescent protein, miRFP720, and the fully NIR Förster resonance energy transfer (FRET) pair miRFP670-miRFP720, which together enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.
Claim 29tool capabilitysupports2018Source 1needs review

miRFP720 and the miRFP670-miRFP720 fully near-infrared FRET pair enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.

Here we report the most red-shifted monomeric near-infrared (NIR) fluorescent protein, miRFP720, and the fully NIR Förster resonance energy transfer (FRET) pair miRFP670-miRFP720, which together enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.
Claim 30tool capabilitysupports2018Source 1needs review

miRFP720 and the miRFP670-miRFP720 fully near-infrared FRET pair enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.

Here we report the most red-shifted monomeric near-infrared (NIR) fluorescent protein, miRFP720, and the fully NIR Förster resonance energy transfer (FRET) pair miRFP670-miRFP720, which together enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.
Claim 31tool capabilitysupports2018Source 1needs review

miRFP720 and the miRFP670-miRFP720 fully near-infrared FRET pair enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.

Here we report the most red-shifted monomeric near-infrared (NIR) fluorescent protein, miRFP720, and the fully NIR Förster resonance energy transfer (FRET) pair miRFP670-miRFP720, which together enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.
Claim 32tool capabilitysupports2018Source 1needs review

miRFP720 and the miRFP670-miRFP720 fully near-infrared FRET pair enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.

Here we report the most red-shifted monomeric near-infrared (NIR) fluorescent protein, miRFP720, and the fully NIR Förster resonance energy transfer (FRET) pair miRFP670-miRFP720, which together enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.
Claim 33tool capabilitysupports2018Source 1needs review

miRFP720 and the miRFP670-miRFP720 fully near-infrared FRET pair enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.

Here we report the most red-shifted monomeric near-infrared (NIR) fluorescent protein, miRFP720, and the fully NIR Förster resonance energy transfer (FRET) pair miRFP670-miRFP720, which together enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.
Claim 34tool capabilitysupports2018Source 1needs review

miRFP720 and the miRFP670-miRFP720 fully near-infrared FRET pair enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.

Here we report the most red-shifted monomeric near-infrared (NIR) fluorescent protein, miRFP720, and the fully NIR Förster resonance energy transfer (FRET) pair miRFP670-miRFP720, which together enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.
Claim 35tool capabilitysupports2018Source 1needs review

miRFP720 and the miRFP670-miRFP720 fully near-infrared FRET pair enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.

Here we report the most red-shifted monomeric near-infrared (NIR) fluorescent protein, miRFP720, and the fully NIR Förster resonance energy transfer (FRET) pair miRFP670-miRFP720, which together enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.
Claim 36tool capabilitysupports2018Source 1needs review

miRFP720 and the miRFP670-miRFP720 fully near-infrared FRET pair enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.

Here we report the most red-shifted monomeric near-infrared (NIR) fluorescent protein, miRFP720, and the fully NIR Förster resonance energy transfer (FRET) pair miRFP670-miRFP720, which together enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.
Claim 37tool developmentsupports2018Source 1needs review

The authors developed a near-infrared biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.

We developed a NIR biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.
Claim 38tool developmentsupports2018Source 1needs review

The authors developed a near-infrared biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.

We developed a NIR biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.
Claim 39tool developmentsupports2018Source 1needs review

The authors developed a near-infrared biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.

We developed a NIR biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.
Claim 40tool developmentsupports2018Source 1needs review

The authors developed a near-infrared biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.

We developed a NIR biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.
Claim 41tool developmentsupports2018Source 1needs review

The authors developed a near-infrared biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.

We developed a NIR biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.
Claim 42tool developmentsupports2018Source 1needs review

The authors developed a near-infrared biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.

We developed a NIR biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.
Claim 43tool developmentsupports2018Source 1needs review

The authors developed a near-infrared biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.

We developed a NIR biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.
Claim 44tool developmentsupports2018Source 1needs review

The authors developed a near-infrared biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.

We developed a NIR biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.
Claim 45tool developmentsupports2018Source 1needs review

The authors developed a near-infrared biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.

We developed a NIR biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways.

Approval Evidence

1 source1 linked approval claimfirst-pass slug mirfp670-mirfp720-fret-pair
the fully NIR Förster resonance energy transfer (FRET) pair miRFP670-miRFP720, which together enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools

Source:

tool capabilitysupports

miRFP720 and the miRFP670-miRFP720 fully near-infrared FRET pair enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.

Here we report the most red-shifted monomeric near-infrared (NIR) fluorescent protein, miRFP720, and the fully NIR Förster resonance energy transfer (FRET) pair miRFP670-miRFP720, which together enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools.

Source:

Comparisons

Source-backed strengths

A key strength is its fully near-infrared spectral placement, which enabled biosensor designs compatible with CFP-YFP imaging and blue-green optogenetic tools. In application, the multiplexed setup allowed direct observation and quantification of ROCK-dependent antagonism between RhoA and Rac1, and simultaneous monitoring of Rac1 activity during optogenetic perturbation.

Source:

fully near-infrared FRET pair

Source:

enabled biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools

Ranked Citations

  1. 1.
    StructuralSource 1Nature Chemical Biology2018Claim 1Claim 2Claim 3

    Extracted from this source document.