Source 1primary paper2020ACS Synthetic Biology
Toolkit/SPLIT
SPLIT
Also known as: Stable Protein Coacervation Using a Light Induced Transition
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
SPLIT (Stable Protein Coacervation Using a Light Induced Transition) is a light-activated multi-component switch engineered to assemble synthetic membraneless organelles. The reported fusion protein combines maltose-binding protein, PhoCl, and two RGG domains so that light triggers a transition to an RGG-driven coacervated state.
Usefulness & Problems
Why this is useful
This system provides optical control over formation of synthetic membraneless organelles using a single light pulse. It is useful for studies that require inducible intracellular coacervation and tunable assembly of protein condensates in living cells.
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
SPLIT addresses the problem of triggering protein coacervation on demand rather than relying on constitutive condensate formation. The reported design enables light-induced assembly of synthetic membraneless organelles in Saccharomyces cerevisiae.
Source:
An optimized version of this system displayed light-induced coacervation in Saccharomyces cerevisiae.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Techniques
No technique tags yet.
Target processes
recombinationInput: Light
Implementation Constraints
The reported construct contains maltose-binding protein as a solubilizing domain, PhoCl as the light-responsive module, and two copies of an RGG domain as the coacervation-driving elements. The evidence indicates activation by a single pulse of light, but the supplied claims do not provide additional construct design parameters, expression details, or delivery guidance.
The supplied evidence does not report quantitative performance metrics, reversibility, kinetics, or off-target effects. Validation is only described for Saccharomyces cerevisiae, and the evidence provided here does not establish independent replication or broader organismal portability.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
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
Approval Evidence
1 source5 linked approval claimsfirst-pass slug split
SPLIT: Stable Protein Coacervation Using a Light Induced Transition
Source:
application resultsupports
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.
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:
engineering resultsupports
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.
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 system was reported to support light-induced coacervation in Saccharomyces cerevisiae, providing a cellular validation of the design. Its construct architecture explicitly couples a solubilizing maltose-binding protein domain, PhoCl, and two RGG domains, and the authors described the resulting organelle assembly as tunable and inducible by a single pulse of light.
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
- 1.