Source 1primary paper2020ACS Sensors
Toolkit/AKAR3EV
AKAR3EV
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
AKAR3EV is a previously developed genetically encoded Förster resonance energy transfer (FRET) biosensor for protein kinase A activity that comprises a CFP/YFP fluorescent protein pair. In the cited 2020 ACS Sensors study, it served as the benchmark comparator for the red-shifted Booster-PKA biosensor.
Usefulness & Problems
Why this is useful
AKAR3EV is useful as a genetically encoded reference-standard PKA activity biosensor within the CFP/YFP FRET class. In the supplied evidence, its main demonstrated utility is as the comparator against which Booster-PKA performance was judged.
Problem solved
AKAR3EV addresses the need to monitor protein kinase A signaling with a genetically encoded FRET biosensor. In the provided study context, it also solves the practical need for an established benchmark for evaluating new PKA biosensor designs.
Problem links
provides a previously developed CFP/YFP-based PKA FRET biosensor comparator
LiteratureAKAR3EV provides a reference standard for PKA FRET biosensing. It gives the authors a benchmark against which to compare the Booster-PKA design.
Source:
AKAR3EV provides a reference standard for PKA FRET biosensing. It gives the authors a benchmark against which to compare the Booster-PKA design.
Published Workflows
Objective: Develop a red-shifted genetically encoded FRET biosensor backbone that avoids the multiplexing and blue-light compatibility limitations of CFP/YFP-based biosensors, then demonstrate its utility in vitro and in vivo.
Why it works: The workflow pairs a favorable red-shifted donor/acceptor set selected by Förster distance calculations with biosensor architecture optimization, then tests whether the resulting design retains biosensor performance while reducing spectral conflicts with other FRET sensors and blue-light optogenetic tools.
Förster resonance energy transfer-based reporting of signaling activityspectral separation to enable multiplexed imaging and optogenetic compatibilityFörster distance-based donor/acceptor selectionoptimization of fluorescent protein orderoptimization of modulatory domain arrangementproof-of-concept application testing
Stages
- 1.Donor-acceptor pair selection by Förster distance calculation(in_silico_filter)
This stage identifies a donor/acceptor pair suitable for building a red-shifted FRET biosensor.
Selection: Favorable donor and acceptor pair chosen by calculating the Förster distance.
- 2.Biosensor backbone optimization(library_design)
This stage converts the selected fluorescent protein pair into a working biosensor backbone.
Selection: Optimization of fluorescent protein order and modulatory domains.
- 3.Benchmarking with a PKA biosensor implementation(functional_characterization)
This stage checks whether the red-shifted backbone retains useful biosensor performance after engineering.
Selection: Comparison of Booster-PKA performance to AKAR3EV.
- 4.Proof-of-concept compatibility demonstrations(confirmatory_validation)
This stage confirms that the engineered spectral shift solves the intended compatibility problems in live-cell use cases.
Selection: Demonstration of simultaneous kinase monitoring and compatibility with a blue light-responsive optogenetic tool.
- 5.In vivo tissue imaging in transgenic mice(in_vivo_validation)
This stage validates that the biosensor can function in living tissues in an animal context, extending beyond in vitro demonstrations.
Selection: Presentation of PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Steps
- 1.Calculate Förster distance to choose donor and acceptor fluorescent proteins
Identify a favorable red-shifted donor/acceptor pair for the biosensor.
Pair selection is performed before biosensor backbone optimization because the fluorescent proteins define the core FRET pair used in the design.
- 2.Optimize fluorescent protein order and modulatory domains to build the Booster backboneengineered biosensor backbone
Convert the selected fluorescent protein pair into a functional red-shifted FRET biosensor backbone.
Architecture optimization follows pair selection because the chosen donor and acceptor must be arranged with modulatory domains to create a working biosensor.
- 3.Implement the Booster backbone as a PKA biosensor and compare it with AKAR3EVbiosensor under test and benchmark comparator
Determine whether the engineered red-shifted backbone retains practical biosensor performance.
Benchmarking occurs after backbone construction to verify that the redesigned sensor remains functionally comparable to an established PKA biosensor before broader application claims.
- 4.Test simultaneous monitoring with a CFP/YFP-based ERK FRET biosensorbiosensor under application test
Demonstrate multiplexed kinase activity imaging with a standard CFP/YFP-based FRET biosensor.
This proof-of-concept follows benchmarking because the authors next test whether the red-shifted design solves the intended multiplexing limitation.
- 5.Test monitoring of PKA activation driven by Beggiatoa photoactivated adenylyl cyclasebiosensor-actuator compatibility pair
Demonstrate compatibility of the red-shifted biosensor with a blue light-responsive optogenetic tool.
After showing multiplexed biosensor compatibility, the authors next test the second intended use case: compatibility with blue light-responsive optogenetic control.
- 6.Image PKA activity in living tissues of transgenic mice expressing Booster-PKAbiosensor under in vivo validation
Extend validation from in vitro proof-of-concept experiments to living tissue imaging in an animal model.
In vivo tissue imaging is presented last as a higher-context validation of versatility after in vitro compatibility demonstrations.
Objective: Develop a red-shifted genetically encoded FRET biosensor backbone that avoids the multiplexing and blue-light incompatibility limitations of CFP/YFP-based FRET biosensors, and demonstrate its utility with a PKA biosensor in vitro and in vivo.
Why it works: The workflow first addresses spectral design by selecting a favorable donor-acceptor pair and optimizing biosensor architecture, then tests whether the resulting backbone retains sensing performance while enabling multiplexing and blue-light optogenetic compatibility.
Förster resonance energy transferPKA activity sensingoptogenetic cAMP-driven PKA activationFörster distance-based fluorophore pair selectionoptimization of fluorescent protein orderoptimization of modulatory domain ordercomparative biosensor benchmarkingproof-of-concept application testing
Stages
- 1.Fluorophore pair selection(in_silico_filter)
This stage identifies a donor-acceptor pair suitable for building red-shifted FRET biosensors.
Selection: Calculated Förster distance used to choose a favorable donor and acceptor pair.
- 2.Backbone optimization(library_design)
This stage converts the selected fluorophore pair into a working biosensor backbone.
Selection: Optimization of the order of fluorescent proteins and modulatory domains.
- 3.Comparator performance testing(confirmatory_validation)
This stage checks whether the red-shifted backbone preserves performance relative to an established CFP/YFP PKA biosensor.
Selection: Comparison of Booster-PKA performance to AKAR3EV.
- 4.Multiplexing proof of concept(functional_characterization)
This stage tests whether the red-shifted design enables simultaneous use with standard CFP/YFP biosensors.
Selection: Ability to monitor two protein kinase activities simultaneously with Booster-PKA and a CFP/YFP ERK FRET biosensor.
- 5.Optogenetic compatibility testing(functional_characterization)
This stage tests whether the red-shifted biosensor can operate with a blue-light optogenetic actuator that would conflict with CFP/YFP biosensors.
Selection: Ability to monitor PKA activation driven by Beggiatoa photoactivated adenylyl cyclase.
- 6.In vivo tissue demonstration(in_vivo_validation)
This stage extends validation from in vitro demonstrations to living tissues in transgenic mice.
Selection: Presentation of PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Steps
- 1.Calculate Förster distance to choose donor and acceptor pair
Identify a favorable fluorescent protein pair for a red-shifted FRET biosensor.
Pair selection is done before backbone optimization because the chosen donor and acceptor define the spectral basis of the biosensor.
- 2.Optimize fluorescent protein and modulatory domain order to build Boosterengineered biosensor backbone
Convert the selected fluorophore pair into a functional red-shifted FRET biosensor backbone.
Architecture optimization follows fluorophore selection because domain order must be tuned around the chosen donor-acceptor pair.
- 3.Benchmark Booster-PKA against AKAR3EVbiosensor and comparator
Test whether the red-shifted PKA biosensor preserves performance relative to an established CFP/YFP biosensor.
Comparator benchmarking is performed after backbone construction to confirm that solving spectral compatibility did not compromise sensing performance.
- 4.Test simultaneous kinase monitoring with Booster-PKA and a CFP/YFP ERK biosensorbiosensor under application test
Demonstrate multiplexed monitoring of two kinase activities in the same setting.
This application test follows comparator benchmarking because multiplexing is a key intended advantage once baseline performance is established.
- 5.Monitor PKA activation driven by Beggiatoa photoactivated adenylyl cyclasebiosensor and optogenetic actuator
Demonstrate compatibility of the red-shifted biosensor with a blue-light optogenetic cAMP generator.
This test follows multiplexing proof of concept because blue-light compatibility is another central design goal enabled by spectral red-shifting.
- 6.Present PKA activity in living tissues of transgenic mice expressing Booster-PKAbiosensor under in vivo validation
Demonstrate that the biosensor can report PKA activity in living mouse tissues.
In vivo demonstration is placed after in vitro proof-of-concept tests as a higher-fidelity validation of practical imaging utility.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Techniques
No technique tags yet.
Target processes
signalingImplementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: sensor
The construct is described only as a genetically encoded FRET biosensor comprising CFP and YFP. No additional implementation details such as phosphorylation substrate sequence, phospho-binding domain, linker architecture, expression system, or imaging settings are provided in the supplied evidence.
The provided evidence does not report AKAR3EV dynamic range, kinetics, localization behavior, or validation across cell types. The same source frames CFP/YFP-based FRET biosensors as poorly suited for single-cell multiplexing and incompatible with blue light-responsive optogenetic tools, which are limitations relevant to the comparator class that includes AKAR3EV.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Booster-PKA can be used simultaneously with a CFP/YFP-based ERK FRET biosensor to monitor activities of two protein kinases.
we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP
Booster-PKA can be used simultaneously with a CFP/YFP-based ERK FRET biosensor to monitor activities of two protein kinases.
we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP
Booster-PKA can be used simultaneously with a CFP/YFP-based ERK FRET biosensor to monitor activities of two protein kinases.
we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP
Booster-PKA can be used simultaneously with a CFP/YFP-based ERK FRET biosensor to monitor activities of two protein kinases.
we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP
Booster-PKA can be used simultaneously with a CFP/YFP-based ERK FRET biosensor to monitor activities of two protein kinases.
we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP
Booster-PKA can be used simultaneously with a CFP/YFP-based ERK FRET biosensor to monitor activities of two protein kinases.
we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP
Booster-PKA can be used simultaneously with a CFP/YFP-based ERK FRET biosensor to monitor activities of two protein kinases.
we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP
Booster-PKA can be used simultaneously with a CFP/YFP-based ERK FRET biosensor to monitor activities of two protein kinases.
we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP
Booster-PKA enabled simultaneous monitoring of two protein kinase activities together with a CFP/YFP-based ERK FRET biosensor.
For the proof of concept, we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP.
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Booster-PKA performance was comparable to that of AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV, a previously developed FRET biosensor comprising CFP and YFP.
Booster-PKA can be used simultaneously with a CFP/YFP-based ERK FRET biosensor to monitor two protein kinase activities.
we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP
Booster-PKA can be used simultaneously with a CFP/YFP-based ERK FRET biosensor to monitor two protein kinase activities.
we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP
Booster-PKA can be used simultaneously with a CFP/YFP-based ERK FRET biosensor to monitor two protein kinase activities.
we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP
Booster-PKA can be used simultaneously with a CFP/YFP-based ERK FRET biosensor to monitor two protein kinase activities.
we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP
Booster-PKA can be used simultaneously with a CFP/YFP-based ERK FRET biosensor to monitor two protein kinase activities.
we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP
Booster-PKA can be used simultaneously with a CFP/YFP-based ERK FRET biosensor to monitor two protein kinase activities.
we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP
Booster-PKA can be used simultaneously with a CFP/YFP-based ERK FRET biosensor to monitor two protein kinase activities.
we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP
Booster-PKA enabled monitoring of PKA activation driven by Beggiatoa photoactivated adenylyl cyclase.
we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP
Booster-PKA enabled monitoring of PKA activation driven by Beggiatoa photoactivated adenylyl cyclase.
we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP
Booster-PKA enabled monitoring of PKA activation driven by Beggiatoa photoactivated adenylyl cyclase.
we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP
Booster-PKA enabled monitoring of PKA activation driven by Beggiatoa photoactivated adenylyl cyclase.
we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP
Booster-PKA enabled monitoring of PKA activation driven by Beggiatoa photoactivated adenylyl cyclase.
we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP
Booster-PKA enabled monitoring of PKA activation driven by Beggiatoa photoactivated adenylyl cyclase.
we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP
Booster-PKA enabled monitoring of PKA activation driven by Beggiatoa photoactivated adenylyl cyclase.
we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP
Booster-PKA is compatible with blue-light optogenetic activation of cAMP signaling using Beggiatoa photoactivated adenylyl cyclase.
Second, we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP.
Booster-PKA is compatible with blue-light optogenetic activation of cAMP signaling using Beggiatoa photoactivated adenylyl cyclase.
Second, we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP.
Booster-PKA is compatible with blue-light optogenetic activation of cAMP signaling using Beggiatoa photoactivated adenylyl cyclase.
Second, we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP.
Booster-PKA is compatible with blue-light optogenetic activation of cAMP signaling using Beggiatoa photoactivated adenylyl cyclase.
Second, we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP.
Booster-PKA is compatible with blue-light optogenetic activation of cAMP signaling using Beggiatoa photoactivated adenylyl cyclase.
Second, we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP.
Booster-PKA is compatible with blue-light optogenetic activation of cAMP signaling using Beggiatoa photoactivated adenylyl cyclase.
Second, we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP.
Booster-PKA is compatible with blue-light optogenetic activation of cAMP signaling using Beggiatoa photoactivated adenylyl cyclase.
Second, we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP.
Booster-PKA is compatible with blue-light optogenetic activation of cAMP signaling using Beggiatoa photoactivated adenylyl cyclase.
Second, we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP.
Booster-PKA was used to monitor PKA activation driven by Beggiatoa photoactivated adenylyl cyclase.
Second, we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP.
The Booster design selected mKOκ and mKate2 as the donor and acceptor pair based on calculated Förster distance.
We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance.
The Booster design selected mKOκ and mKate2 as the donor and acceptor pair based on calculated Förster distance.
We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance.
The Booster design selected mKOκ and mKate2 as the donor and acceptor pair based on calculated Förster distance.
We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance.
The Booster design selected mKOκ and mKate2 as the donor and acceptor pair based on calculated Förster distance.
We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance.
The Booster design selected mKOκ and mKate2 as the donor and acceptor pair based on calculated Förster distance.
We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance.
The Booster design selected mKOκ and mKate2 as the donor and acceptor pair based on calculated Förster distance.
We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance.
The Booster design selected mKOκ and mKate2 as the donor and acceptor pair based on calculated Förster distance.
We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance.
The Booster design selected mKOκ and mKate2 as the donor and acceptor pair based on calculated Förster distance.
We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance.
Booster is a red-shifted genetically encoded FRET biosensor backbone developed to overcome limitations of CFP/YFP-based FRET biosensors for multiplexing and blue-light optogenetic compatibility.
To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths... we developed a FRET biosensor backbone named "Booster".
Booster is a red-shifted genetically encoded FRET biosensor backbone developed to overcome limitations of CFP/YFP-based FRET biosensors for multiplexing and blue-light optogenetic compatibility.
To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths... we developed a FRET biosensor backbone named "Booster".
Booster is a red-shifted genetically encoded FRET biosensor backbone developed to overcome limitations of CFP/YFP-based FRET biosensors for multiplexing and blue-light optogenetic compatibility.
To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths... we developed a FRET biosensor backbone named "Booster".
Booster is a red-shifted genetically encoded FRET biosensor backbone developed to overcome limitations of CFP/YFP-based FRET biosensors for multiplexing and blue-light optogenetic compatibility.
To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths... we developed a FRET biosensor backbone named "Booster".
Booster is a red-shifted genetically encoded FRET biosensor backbone developed to overcome limitations of CFP/YFP-based FRET biosensors for multiplexing and blue-light optogenetic compatibility.
To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths... we developed a FRET biosensor backbone named "Booster".
Booster is a red-shifted genetically encoded FRET biosensor backbone developed to overcome limitations of CFP/YFP-based FRET biosensors for multiplexing and blue-light optogenetic compatibility.
To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths... we developed a FRET biosensor backbone named "Booster".
Booster is a red-shifted genetically encoded FRET biosensor backbone developed to overcome limitations of CFP/YFP-based FRET biosensors for multiplexing and blue-light optogenetic compatibility.
To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths... we developed a FRET biosensor backbone named "Booster".
Booster is a red-shifted genetically encoded FRET biosensor backbone developed to overcome limitations of CFP/YFP-based FRET biosensors for multiplexing and blue-light optogenetic compatibility.
To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths... we developed a FRET biosensor backbone named "Booster".
The authors developed red-shifted FRET biosensors and a biosensor backbone named Booster by choosing mKOκ and mKate2 and optimizing fluorescent protein order and modulatory domains.
We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance. By optimizing the order of fluorescent proteins and modulatory domains of the FRET biosensors, we developed a FRET biosensor backbone named "Booster".
Booster is a red-shifted genetically encoded FRET biosensor backbone developed by optimizing fluorescent protein order and modulatory domains.
To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths. We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance. By optimizing the order of fluorescent proteins and modulatory domains of the FRET biosensors, we developed a FRET biosensor backbone named "Booster".
Booster is a red-shifted genetically encoded FRET biosensor backbone developed by optimizing fluorescent protein order and modulatory domains.
To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths. We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance. By optimizing the order of fluorescent proteins and modulatory domains of the FRET biosensors, we developed a FRET biosensor backbone named "Booster".
Booster is a red-shifted genetically encoded FRET biosensor backbone developed by optimizing fluorescent protein order and modulatory domains.
To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths. We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance. By optimizing the order of fluorescent proteins and modulatory domains of the FRET biosensors, we developed a FRET biosensor backbone named "Booster".
Booster is a red-shifted genetically encoded FRET biosensor backbone developed by optimizing fluorescent protein order and modulatory domains.
To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths. We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance. By optimizing the order of fluorescent proteins and modulatory domains of the FRET biosensors, we developed a FRET biosensor backbone named "Booster".
Booster is a red-shifted genetically encoded FRET biosensor backbone developed by optimizing fluorescent protein order and modulatory domains.
To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths. We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance. By optimizing the order of fluorescent proteins and modulatory domains of the FRET biosensors, we developed a FRET biosensor backbone named "Booster".
Booster is a red-shifted genetically encoded FRET biosensor backbone developed by optimizing fluorescent protein order and modulatory domains.
To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths. We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance. By optimizing the order of fluorescent proteins and modulatory domains of the FRET biosensors, we developed a FRET biosensor backbone named "Booster".
Booster is a red-shifted genetically encoded FRET biosensor backbone developed by optimizing fluorescent protein order and modulatory domains.
To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths. We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Förster distance. By optimizing the order of fluorescent proteins and modulatory domains of the FRET biosensors, we developed a FRET biosensor backbone named "Booster".
Booster-PKA was used to present PKA activity in living tissues of transgenic mice.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster-PKA was used to present PKA activity in living tissues of transgenic mice.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster-PKA was used to present PKA activity in living tissues of transgenic mice.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster-PKA was used to present PKA activity in living tissues of transgenic mice.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster-PKA was used to present PKA activity in living tissues of transgenic mice.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster-PKA was used to present PKA activity in living tissues of transgenic mice.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster-PKA was used to present PKA activity in living tissues of transgenic mice.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster-PKA was used to present PKA activity in living tissues of transgenic mice.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster-PKA was used to present PKA activity in living tissues of transgenic mice.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster-PKA was used to present PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster-PKA was used to present PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster-PKA was used to present PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster-PKA was used to present PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster-PKA was used to present PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster-PKA was used to present PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster-PKA was used to present PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Booster biosensors are effective and versatile imaging tools in vitro and in vivo.
Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.
Approval Evidence
1 source3 linked approval claimsfirst-pass slug akar3ev
AKAR3EV, a previously developed FRET biosensor comprising CFP and YFP
Source:
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV, a previously developed FRET biosensor comprising CFP and YFP.
Source:
comparative performancesupports
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Source:
comparative performancesupports
Booster-PKA performance was comparable to AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV
Source:
comparative performancesupports
Booster-PKA performance was comparable to that of AKAR3EV.
The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV, a previously developed FRET biosensor comprising CFP and YFP.
Source:
Comparisons
Source-stated alternatives
The paper presents Booster-PKA as a red-shifted alternative with comparable performance.; Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Source:
The paper presents Booster-PKA as a red-shifted alternative with comparable performance.
Source:
Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Source-backed strengths
The supplied evidence identifies AKAR3EV as an established, previously developed PKA FRET biosensor built from CFP and YFP. Booster-PKA was reported to have performance comparable to AKAR3EV, indicating that AKAR3EV represents a meaningful performance standard in this assay context.
Compared with biosensors
Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Shared frame: source-stated alternative in extracted literature
Strengths here: serves as a performance comparator for Booster-PKA; serves as a benchmark comparator for Booster-PKA.
Relative tradeoffs: comprises CFP and YFP; CFP/YFP-based FRET biosensors are described as incompatible with blue-light optogenetic tools; CFP/YFP-based FRET biosensors are described as limiting multiplexed use within a single cell.
Source:
Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Compared with biosensors for active Rho detection
Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Shared frame: source-stated alternative in extracted literature
Strengths here: serves as a performance comparator for Booster-PKA; serves as a benchmark comparator for Booster-PKA.
Relative tradeoffs: comprises CFP and YFP; CFP/YFP-based FRET biosensors are described as incompatible with blue-light optogenetic tools; CFP/YFP-based FRET biosensors are described as limiting multiplexed use within a single cell.
Source:
Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Compared with Booster
The paper presents Booster-PKA as a red-shifted alternative with comparable performance.; Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Shared frame: source-stated alternative in extracted literature
Strengths here: serves as a performance comparator for Booster-PKA; serves as a benchmark comparator for Booster-PKA.
Relative tradeoffs: comprises CFP and YFP; CFP/YFP-based FRET biosensors are described as incompatible with blue-light optogenetic tools; CFP/YFP-based FRET biosensors are described as limiting multiplexed use within a single cell.
Source:
The paper presents Booster-PKA as a red-shifted alternative with comparable performance.
Source:
Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Compared with Booster-PKA
The paper presents Booster-PKA as a red-shifted alternative with comparable performance.; Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Shared frame: source-stated alternative in extracted literature
Strengths here: serves as a performance comparator for Booster-PKA; serves as a benchmark comparator for Booster-PKA.
Relative tradeoffs: comprises CFP and YFP; CFP/YFP-based FRET biosensors are described as incompatible with blue-light optogenetic tools; CFP/YFP-based FRET biosensors are described as limiting multiplexed use within a single cell.
Source:
The paper presents Booster-PKA as a red-shifted alternative with comparable performance.
Source:
Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Compared with fluorescent protein based reporters and biosensors
Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Shared frame: source-stated alternative in extracted literature
Strengths here: serves as a performance comparator for Booster-PKA; serves as a benchmark comparator for Booster-PKA.
Relative tradeoffs: comprises CFP and YFP; CFP/YFP-based FRET biosensors are described as incompatible with blue-light optogenetic tools; CFP/YFP-based FRET biosensors are described as limiting multiplexed use within a single cell.
Source:
Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Compared with FRET
Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Shared frame: source-stated alternative in extracted literature
Strengths here: serves as a performance comparator for Booster-PKA; serves as a benchmark comparator for Booster-PKA.
Relative tradeoffs: comprises CFP and YFP; CFP/YFP-based FRET biosensors are described as incompatible with blue-light optogenetic tools; CFP/YFP-based FRET biosensors are described as limiting multiplexed use within a single cell.
Source:
Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Compared with genetically engineered biosensors
Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Shared frame: source-stated alternative in extracted literature
Strengths here: serves as a performance comparator for Booster-PKA; serves as a benchmark comparator for Booster-PKA.
Relative tradeoffs: comprises CFP and YFP; CFP/YFP-based FRET biosensors are described as incompatible with blue-light optogenetic tools; CFP/YFP-based FRET biosensors are described as limiting multiplexed use within a single cell.
Source:
Booster-PKA is the direct alternative presented in this paper. The abstract also refers more broadly to red-shifted Booster biosensors as an alternative to CFP/YFP-based FRET biosensors.
Ranked Citations
- 1.