Source 1review2020Current Opinion in Cell Biology
Toolkit/PhoCl
PhoCl
Also known as: photocleavable protein
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
PhoCl is a light-responsive protein domain that cleaves upon 405 nm illumination. In the SPLIT system, it was fused between maltose-binding protein and a tandem RGG coacervation module to trigger light-induced assembly of synthetic membraneless organelles in Saccharomyces cerevisiae after a single light pulse.
Usefulness & Problems
Why this is useful
PhoCl is useful as an optogenetic trigger for irreversible protein-state changes because its light response is encoded directly in a protein domain that undergoes cleavage. In the cited application, this enabled single-pulse control over coacervation and the formation of tunable synthetic membraneless organelles.
Source:
An optimized version of this system displayed light-induced coacervation in Saccharomyces cerevisiae.
Source:
Several seconds of illumination at 405 nm is sufficient to cleave PhoCl, removing the solubilization domain and enabling RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
Problem solved
This tool helps solve the problem of how to convert a brief light input into assembly of a coacervating protein system. In the reported design, PhoCl-mediated cleavage removed a solubilizing constraint from a fusion protein, enabling RGG-driven coacervation in yeast.
Source:
An optimized version of this system displayed light-induced coacervation in Saccharomyces cerevisiae.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level protein part used inside a larger architecture that realizes a mechanism.
Mechanisms
PhotocleavageTechniques
No technique tags yet.
Target processes
No target processes tagged yet.
Input: Light
Implementation Constraints
In the cited construct, PhoCl was incorporated into a fusion protein containing a solubilizing maltose-binding protein domain and two copies of an RGG domain. Activation was achieved with 405 nm light, and the demonstrated application was in Saccharomyces cerevisiae.
The supplied evidence supports PhoCl only in the context of the SPLIT fusion construct and does not provide standalone performance metrics such as cleavage efficiency, kinetics, dynamic range, or phototoxicity. Validation in the provided evidence is limited to a yeast coacervation application.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Source 2primary paper2020ACS Synthetic Biology
Source 3review2021Advanced Biology
Ranked Claims
An optimized version of the system displayed light-induced coacervation in Saccharomyces cerevisiae.
An optimized version of this system displayed light-induced coacervation in Saccharomyces cerevisiae.
An optimized version of the system displayed light-induced coacervation in Saccharomyces cerevisiae.
An optimized version of this system displayed light-induced coacervation in Saccharomyces cerevisiae.
An optimized version of the system displayed light-induced coacervation in Saccharomyces cerevisiae.
An optimized version of this system displayed light-induced coacervation in Saccharomyces cerevisiae.
An optimized version of the system displayed light-induced coacervation in Saccharomyces cerevisiae.
An optimized version of this system displayed light-induced coacervation in Saccharomyces cerevisiae.
An optimized version of the system displayed light-induced coacervation in Saccharomyces cerevisiae.
An optimized version of this system displayed light-induced coacervation in Saccharomyces cerevisiae.
An optimized version of the system displayed light-induced coacervation in Saccharomyces cerevisiae.
An optimized version of this system displayed light-induced coacervation in Saccharomyces cerevisiae.
An optimized version of the system displayed light-induced coacervation in Saccharomyces cerevisiae.
An optimized version of this system displayed light-induced coacervation in Saccharomyces cerevisiae.
The fusion protein contains a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
We developed a fusion protein containing a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
The fusion protein contains a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
We developed a fusion protein containing a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
The fusion protein contains a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
We developed a fusion protein containing a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
The fusion protein contains a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
We developed a fusion protein containing a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
The fusion protein contains a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
We developed a fusion protein containing a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
The fusion protein contains a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
We developed a fusion protein containing a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
The fusion protein contains a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
We developed a fusion protein containing a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
The authors engineered a coacervating protein to create tunable synthetic membraneless organelles that assemble in response to a single pulse of light.
We have engineered a coacervating protein to create tunable, synthetic membraneless organelles that assemble in response to a single pulse of light.
The authors engineered a coacervating protein to create tunable synthetic membraneless organelles that assemble in response to a single pulse of light.
We have engineered a coacervating protein to create tunable, synthetic membraneless organelles that assemble in response to a single pulse of light.
The authors engineered a coacervating protein to create tunable synthetic membraneless organelles that assemble in response to a single pulse of light.
We have engineered a coacervating protein to create tunable, synthetic membraneless organelles that assemble in response to a single pulse of light.
The authors engineered a coacervating protein to create tunable synthetic membraneless organelles that assemble in response to a single pulse of light.
We have engineered a coacervating protein to create tunable, synthetic membraneless organelles that assemble in response to a single pulse of light.
The authors engineered a coacervating protein to create tunable synthetic membraneless organelles that assemble in response to a single pulse of light.
We have engineered a coacervating protein to create tunable, synthetic membraneless organelles that assemble in response to a single pulse of light.
The authors engineered a coacervating protein to create tunable synthetic membraneless organelles that assemble in response to a single pulse of light.
We have engineered a coacervating protein to create tunable, synthetic membraneless organelles that assemble in response to a single pulse of light.
The authors engineered a coacervating protein to create tunable synthetic membraneless organelles that assemble in response to a single pulse of light.
We have engineered a coacervating protein to create tunable, synthetic membraneless organelles that assemble in response to a single pulse of light.
Several seconds of 405 nm illumination is sufficient to cleave PhoCl, remove the solubilization domain, and enable RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
Several seconds of illumination at 405 nm is sufficient to cleave PhoCl, removing the solubilization domain and enabling RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
coacervation time within minutesillumination duration several secondsillumination wavelength 405 nm
Several seconds of 405 nm illumination is sufficient to cleave PhoCl, remove the solubilization domain, and enable RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
Several seconds of illumination at 405 nm is sufficient to cleave PhoCl, removing the solubilization domain and enabling RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
coacervation time within minutesillumination duration several secondsillumination wavelength 405 nm
Several seconds of 405 nm illumination is sufficient to cleave PhoCl, remove the solubilization domain, and enable RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
Several seconds of illumination at 405 nm is sufficient to cleave PhoCl, removing the solubilization domain and enabling RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
coacervation time within minutesillumination duration several secondsillumination wavelength 405 nm
Several seconds of 405 nm illumination is sufficient to cleave PhoCl, remove the solubilization domain, and enable RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
Several seconds of illumination at 405 nm is sufficient to cleave PhoCl, removing the solubilization domain and enabling RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
coacervation time within minutesillumination duration several secondsillumination wavelength 405 nm
Several seconds of 405 nm illumination is sufficient to cleave PhoCl, remove the solubilization domain, and enable RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
Several seconds of illumination at 405 nm is sufficient to cleave PhoCl, removing the solubilization domain and enabling RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
coacervation time within minutesillumination duration several secondsillumination wavelength 405 nm
Several seconds of 405 nm illumination is sufficient to cleave PhoCl, remove the solubilization domain, and enable RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
Several seconds of illumination at 405 nm is sufficient to cleave PhoCl, removing the solubilization domain and enabling RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
coacervation time within minutesillumination duration several secondsillumination wavelength 405 nm
Several seconds of 405 nm illumination is sufficient to cleave PhoCl, remove the solubilization domain, and enable RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
Several seconds of illumination at 405 nm is sufficient to cleave PhoCl, removing the solubilization domain and enabling RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
coacervation time within minutesillumination duration several secondsillumination wavelength 405 nm
In the reported system, coacervation is driven by the LAF-1 RGG domain and light responsiveness is provided by PhoCl cleavage in response to 405 nm light.
Coacervation is driven by the intrinsically disordered RGG domain from the protein LAF-1, and opto-responsiveness is coded by the protein PhoCl, which cleaves in response to 405 nm light.
activation wavelength 405 nm
In the reported system, coacervation is driven by the LAF-1 RGG domain and light responsiveness is provided by PhoCl cleavage in response to 405 nm light.
Coacervation is driven by the intrinsically disordered RGG domain from the protein LAF-1, and opto-responsiveness is coded by the protein PhoCl, which cleaves in response to 405 nm light.
activation wavelength 405 nm
In the reported system, coacervation is driven by the LAF-1 RGG domain and light responsiveness is provided by PhoCl cleavage in response to 405 nm light.
Coacervation is driven by the intrinsically disordered RGG domain from the protein LAF-1, and opto-responsiveness is coded by the protein PhoCl, which cleaves in response to 405 nm light.
activation wavelength 405 nm
In the reported system, coacervation is driven by the LAF-1 RGG domain and light responsiveness is provided by PhoCl cleavage in response to 405 nm light.
Coacervation is driven by the intrinsically disordered RGG domain from the protein LAF-1, and opto-responsiveness is coded by the protein PhoCl, which cleaves in response to 405 nm light.
activation wavelength 405 nm
In the reported system, coacervation is driven by the LAF-1 RGG domain and light responsiveness is provided by PhoCl cleavage in response to 405 nm light.
Coacervation is driven by the intrinsically disordered RGG domain from the protein LAF-1, and opto-responsiveness is coded by the protein PhoCl, which cleaves in response to 405 nm light.
activation wavelength 405 nm
In the reported system, coacervation is driven by the LAF-1 RGG domain and light responsiveness is provided by PhoCl cleavage in response to 405 nm light.
Coacervation is driven by the intrinsically disordered RGG domain from the protein LAF-1, and opto-responsiveness is coded by the protein PhoCl, which cleaves in response to 405 nm light.
activation wavelength 405 nm
In the reported system, coacervation is driven by the LAF-1 RGG domain and light responsiveness is provided by PhoCl cleavage in response to 405 nm light.
Coacervation is driven by the intrinsically disordered RGG domain from the protein LAF-1, and opto-responsiveness is coded by the protein PhoCl, which cleaves in response to 405 nm light.
activation wavelength 405 nm
Genetically encoded fluorescent biosensors and optogenetic actuators form an extensive molecular toolkit for monitoring and manipulating signaling activities with high spatiotemporal precision.
The review covers basic concepts and recent advances in the development and application of genetically encodable biosensors and optogenetic tools for understanding signaling activity.
Fluorescent biosensors are used to monitor signaling activities.
Optogenetic actuators are used to manipulate signaling activities.
Approval Evidence
3 sources3 linked approval claimsfirst-pass slug phocl
The web research summary states that the review explicitly discusses PhoCl as a photocleavable protein and a tool/component in the anchor review.
Source:
web_research_summary lists PhoCl as an explicitly supported tool mentioned in the anchor review and describes it as a photocleavable optogenetic protein for controlling localization and enzyme activity
Source:
opto-responsiveness is coded by the protein PhoCl, which cleaves in response to 405 nm light
Source:
construct compositionsupports
The fusion protein contains a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
We developed a fusion protein containing a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain.
Source:
functional performancesupports
Several seconds of 405 nm illumination is sufficient to cleave PhoCl, remove the solubilization domain, and enable RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
Several seconds of illumination at 405 nm is sufficient to cleave PhoCl, removing the solubilization domain and enabling RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions.
Source:
mechanismsupports
In the reported system, coacervation is driven by the LAF-1 RGG domain and light responsiveness is provided by PhoCl cleavage in response to 405 nm light.
Coacervation is driven by the intrinsically disordered RGG domain from the protein LAF-1, and opto-responsiveness is coded by the protein PhoCl, which cleaves in response to 405 nm light.
Source:
Comparisons
Source-backed strengths
The cited study reports that an optimized system displayed light-induced coacervation in Saccharomyces cerevisiae. The response was triggered by a single 405 nm light pulse, supporting its use for temporally precise induction of synthetic membraneless organelle assembly.
Source:
We have engineered a coacervating protein to create tunable, synthetic membraneless organelles that assemble in response to a single pulse of light.
Ranked Citations
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