Source 1primary paper2018Nature Chemical Biology
Toolkit/CFP-YFP FRET biosensors
CFP-YFP FRET biosensors
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
CFP-YFP FRET biosensors are genetically encoded visible-range reporter constructs used in the cited study to measure RhoA activity and Rac1-GDI binding. In that work, they were combined with a near-infrared Rac1 biosensor to enable parallel imaging of Rho GTPase signaling during optogenetic manipulation of Rac1.
Usefulness & Problems
Why this is useful
These biosensors provide established CFP-YFP readouts for RhoA signaling state and Rac1-GDI interaction state in live-cell imaging experiments. In the cited study, their main utility was as complementary visible-spectrum channels that could be paired with a near-infrared Rac1 biosensor for multiplex observation of coordinated Rho GTPase regulation.
Source:
These CFP-YFP FRET biosensors report RhoA activity and Rac1-GDI binding. In this paper they serve as parallel readouts combined with the NIR Rac1 biosensor.
Source:
monitoring RhoA signaling
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monitoring Rac1-GDI binding
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multiplex imaging with NIR biosensors
Problem solved
They help solve the need for simultaneous measurement of multiple Rho GTPase-related signals by supplying visible-range FRET reporters for RhoA and Rac1-GDI binding. In the reported application, this enabled direct observation of Rac1 activity together with additional pathway readouts during Rac1 optogenetic perturbation.
Source:
They provide established visible-range pathway readouts that can be paired with the new NIR biosensor for multiplex measurements.
Source:
providing parallel visible-spectrum readouts for multiplex pathway imaging
Problem links
providing parallel visible-spectrum readouts for multiplex pathway imaging
LiteratureThey provide established visible-range pathway readouts that can be paired with the new NIR biosensor for multiplex measurements.
Source:
They provide established visible-range pathway readouts that can be paired with the new NIR biosensor for multiplex measurements.
Published Workflows
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 reusable architecture pattern for arranging parts into an engineered system.
Mechanisms
biosensing of protein activity statebiosensing of protein-protein bindingförster resonance energy transferTechniques
No technique tags yet.
Target processes
No target processes tagged yet.
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: sensor
Use in the cited work required expression of CFP-YFP FRET biosensor constructs for RhoA and Rac1-GDI binding together with compatible fluorescence imaging channels. The evidence supports combination with a near-infrared Rac1 biosensor in a multiplex imaging setup, but it does not provide further construct design or delivery details.
The supplied evidence does not describe the detailed construct architecture, dynamic range, kinetics, or calibration of the CFP-YFP biosensors in this study. The source context also indicates that visible-range probes alone do not resolve spectral-overlap constraints when combined with blue-green optogenetic tools.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.
and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.
and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.
and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.
and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.
and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.
and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.
and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.
and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
The authors simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.
and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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 source2 linked approval claimsfirst-pass slug cfp-yfp-fret-biosensors
we combined the Rac1 biosensor with CFP-YFP FRET biosensors for RhoA and for Rac1-GDI binding
Source:
biological observationsupports
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
Source:
biological observationsupports
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
Source:
Comparisons
Source-stated alternatives
The abstract presents the NIR miRFP670-miRFP720-based biosensor strategy as a complementary spectral alternative.
Source:
The abstract presents the NIR miRFP670-miRFP720-based biosensor strategy as a complementary spectral alternative.
Source-backed strengths
The study used these CFP-YFP biosensors in a multiplexed imaging configuration with a near-infrared Rac1 biosensor, demonstrating parallel monitoring of RhoA activity, Rac1 activity, and Rac1-GDI binding. This setup supported biological observations that Rac1 activity and GDI binding depend on their spatiotemporal coordination and that RhoA-Rac1 antagonism depends on the RhoA effector ROCK.
Source:
used in combination with the NIR Rac1 biosensor for concurrent measurements
Compared with AKAR3EV
CFP-YFP FRET biosensors and AKAR3EV address a similar problem space.
Shared frame: same top-level item type; shared mechanisms: förster resonance energy transfer
Compared with joining proteins in creative ways
CFP-YFP FRET biosensors and joining proteins in creative ways address a similar problem space.
Shared frame: same top-level item type
Compared with tethered PEs
CFP-YFP FRET biosensors and tethered PEs address a similar problem space.
Shared frame: same top-level item type
Ranked Citations
- 1.