Source 1primary paper2016DukeSpace (Duke University)
Toolkit/Rho1-CRY2 fusion construct
Rho1-CRY2 fusion construct
Also known as: Rho variants fused to CRY2
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The Rho1-CRY2 fusion construct is a proposed optogenetic multi-component switch for Drosophila in which a small G protein Rho variant would be fused to CRY2. The intended function is blue-light-dependent recruitment of Rho1-CRY2 to membrane-anchored CIB to control subcellular localization and downstream events, but the available evidence indicates the construct was still being cloned rather than functionally validated.
Usefulness & Problems
Why this is useful
This design is intended to provide spatial and temporal control of protein localization in Drosophila using light. The evidence supports its conceptual utility for recruiting a Rho fusion to membrane-anchored CIB, but does not document successful deployment or biological outcomes.
Problem solved
The construct is meant to address the problem of controlling localization of a Rho-family small G protein with high spatial and temporal precision in Drosophila. The supplied evidence only supports that this was an intended application of adapting the CRY2/CIB system to Drosophila-specific vectors.
Problem links
Need inducible protein relocalization or recruitment
DerivedThe Rho1-CRY2 fusion construct is a proposed optogenetic multi-component switch in which a small G protein Rho variant is fused to CRY2 for light-dependent control in Drosophila. The intended design is that blue light would recruit Rho1-CRY2 to membrane-anchored CIB, enabling spatial and temporal control of protein localization and downstream events, but the provided evidence indicates this construct was still at the cloning stage.
Need precise spatiotemporal control with light input
DerivedThe Rho1-CRY2 fusion construct is a proposed optogenetic multi-component switch in which a small G protein Rho variant is fused to CRY2 for light-dependent control in Drosophila. The intended design is that blue light would recruit Rho1-CRY2 to membrane-anchored CIB, enabling spatial and temporal control of protein localization and downstream events, but the provided evidence indicates this construct was still at the cloning stage.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
light-controlled subcellular recruitmentlight-controlled subcellular recruitmentlight-induced heterodimerizationlight-induced heterodimerizationTechniques
No technique tags yet.
Target processes
localizationInput: Light
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: multi component delivery burdenimplementation constraint: spectral hardware requirementoperating role: actuatoroperating role: regulatorswitch architecture: multi componentswitch architecture: recruitment
The available evidence indicates that variants of the small G protein Rho were being cloned as fusion proteins with CRY2. The intended implementation also requires membrane-anchored CIB and adaptation of CRY2/CIB constructs into Drosophila-specific vectors; no further construct architecture, expression conditions, or cofactor requirements are described in the supplied evidence.
The main limitation is that the Rho1-CRY2 construct appears to have remained at the cloning stage in the cited evidence. There is no reported demonstration of expression, light-induced membrane recruitment, downstream signaling control, or phenotypic validation for this specific fusion.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
The authors attempted to adapt the CRY2/CIB system to Drosophila using CRY2 and CIB constructs in Drosophila-specific vectors.
Using cloning techniques to generate CRY2 and CIB constructs in Drosophila specific vectors, we have attempted to adapt the CRY2/CIB system to Drosophila.
The authors attempted to adapt the CRY2/CIB system to Drosophila using CRY2 and CIB constructs in Drosophila-specific vectors.
Using cloning techniques to generate CRY2 and CIB constructs in Drosophila specific vectors, we have attempted to adapt the CRY2/CIB system to Drosophila.
The authors attempted to adapt the CRY2/CIB system to Drosophila using CRY2 and CIB constructs in Drosophila-specific vectors.
Using cloning techniques to generate CRY2 and CIB constructs in Drosophila specific vectors, we have attempted to adapt the CRY2/CIB system to Drosophila.
The authors attempted to adapt the CRY2/CIB system to Drosophila using CRY2 and CIB constructs in Drosophila-specific vectors.
Using cloning techniques to generate CRY2 and CIB constructs in Drosophila specific vectors, we have attempted to adapt the CRY2/CIB system to Drosophila.
The authors attempted to adapt the CRY2/CIB system to Drosophila using CRY2 and CIB constructs in Drosophila-specific vectors.
Using cloning techniques to generate CRY2 and CIB constructs in Drosophila specific vectors, we have attempted to adapt the CRY2/CIB system to Drosophila.
An oligomerizing version of CRY2 was tested as a tool for negative regulation of targeted proteins in Drosophila.
We tested an oligomerizing version of the CRY2 component as a tool for the negative regulation of targeted proteins in Drosophila.
An oligomerizing version of CRY2 was tested as a tool for negative regulation of targeted proteins in Drosophila.
We tested an oligomerizing version of the CRY2 component as a tool for the negative regulation of targeted proteins in Drosophila.
An oligomerizing version of CRY2 was tested as a tool for negative regulation of targeted proteins in Drosophila.
We tested an oligomerizing version of the CRY2 component as a tool for the negative regulation of targeted proteins in Drosophila.
An oligomerizing version of CRY2 was tested as a tool for negative regulation of targeted proteins in Drosophila.
We tested an oligomerizing version of the CRY2 component as a tool for the negative regulation of targeted proteins in Drosophila.
An oligomerizing version of CRY2 was tested as a tool for negative regulation of targeted proteins in Drosophila.
We tested an oligomerizing version of the CRY2 component as a tool for the negative regulation of targeted proteins in Drosophila.
An oligomerizing version of CRY2 was tested as a tool for negative regulation of targeted proteins in Drosophila.
We tested an oligomerizing version of the CRY2 component as a tool for the negative regulation of targeted proteins in Drosophila.
An oligomerizing version of CRY2 was tested as a tool for negative regulation of targeted proteins in Drosophila.
We tested an oligomerizing version of the CRY2 component as a tool for the negative regulation of targeted proteins in Drosophila.
An oligomerizing version of CRY2 was tested as a tool for negative regulation of targeted proteins in Drosophila.
We tested an oligomerizing version of the CRY2 component as a tool for the negative regulation of targeted proteins in Drosophila.
An oligomerizing version of CRY2 was tested as a tool for negative regulation of targeted proteins in Drosophila.
We tested an oligomerizing version of the CRY2 component as a tool for the negative regulation of targeted proteins in Drosophila.
An oligomerizing version of CRY2 was tested as a tool for negative regulation of targeted proteins in Drosophila.
We tested an oligomerizing version of the CRY2 component as a tool for the negative regulation of targeted proteins in Drosophila.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
The authors were unable to repeat in Drosophila the clustering results previously observed in yeast for the oligomerizing CRY2 component.
Although we were unable to repeat the clustering results observed in yeast
The authors were unable to repeat in Drosophila the clustering results previously observed in yeast for the oligomerizing CRY2 component.
Although we were unable to repeat the clustering results observed in yeast
The authors were unable to repeat in Drosophila the clustering results previously observed in yeast for the oligomerizing CRY2 component.
Although we were unable to repeat the clustering results observed in yeast
The authors were unable to repeat in Drosophila the clustering results previously observed in yeast for the oligomerizing CRY2 component.
Although we were unable to repeat the clustering results observed in yeast
The authors were unable to repeat in Drosophila the clustering results previously observed in yeast for the oligomerizing CRY2 component.
Although we were unable to repeat the clustering results observed in yeast
The authors were unable to repeat in Drosophila the clustering results previously observed in yeast for the oligomerizing CRY2 component.
Although we were unable to repeat the clustering results observed in yeast
The authors were unable to repeat in Drosophila the clustering results previously observed in yeast for the oligomerizing CRY2 component.
Although we were unable to repeat the clustering results observed in yeast
The authors were unable to repeat in Drosophila the clustering results previously observed in yeast for the oligomerizing CRY2 component.
Although we were unable to repeat the clustering results observed in yeast
The authors were unable to repeat in Drosophila the clustering results previously observed in yeast for the oligomerizing CRY2 component.
Although we were unable to repeat the clustering results observed in yeast
The authors were unable to repeat in Drosophila the clustering results previously observed in yeast for the oligomerizing CRY2 component.
Although we were unable to repeat the clustering results observed in yeast
The light activation protocol was sensitive to inadvertent light stimulation during preparation for imaging.
discovered the sensitivity of the system to inadvertent light stimulation during preparation for imaging
The light activation protocol was sensitive to inadvertent light stimulation during preparation for imaging.
discovered the sensitivity of the system to inadvertent light stimulation during preparation for imaging
The light activation protocol was sensitive to inadvertent light stimulation during preparation for imaging.
discovered the sensitivity of the system to inadvertent light stimulation during preparation for imaging
The light activation protocol was sensitive to inadvertent light stimulation during preparation for imaging.
discovered the sensitivity of the system to inadvertent light stimulation during preparation for imaging
The light activation protocol was sensitive to inadvertent light stimulation during preparation for imaging.
discovered the sensitivity of the system to inadvertent light stimulation during preparation for imaging
Germline transformants of the CIBN component had been generated, while CRY2 germline transformants were still in progress.
Thus far, germline transformants of the CIBN component have been generated, and work will continue to generate the CRY2 germline transformants.
Germline transformants of the CIBN component had been generated, while CRY2 germline transformants were still in progress.
Thus far, germline transformants of the CIBN component have been generated, and work will continue to generate the CRY2 germline transformants.
Germline transformants of the CIBN component had been generated, while CRY2 germline transformants were still in progress.
Thus far, germline transformants of the CIBN component have been generated, and work will continue to generate the CRY2 germline transformants.
Germline transformants of the CIBN component had been generated, while CRY2 germline transformants were still in progress.
Thus far, germline transformants of the CIBN component have been generated, and work will continue to generate the CRY2 germline transformants.
Germline transformants of the CIBN component had been generated, while CRY2 germline transformants were still in progress.
Thus far, germline transformants of the CIBN component have been generated, and work will continue to generate the CRY2 germline transformants.
Germline transformants of the CIBN component had been generated, while CRY2 germline transformants were still in progress.
Thus far, germline transformants of the CIBN component have been generated, and work will continue to generate the CRY2 germline transformants.
Germline transformants of the CIBN component had been generated, while CRY2 germline transformants were still in progress.
Thus far, germline transformants of the CIBN component have been generated, and work will continue to generate the CRY2 germline transformants.
Germline transformants of the CIBN component had been generated, while CRY2 germline transformants were still in progress.
Thus far, germline transformants of the CIBN component have been generated, and work will continue to generate the CRY2 germline transformants.
Germline transformants of the CIBN component had been generated, while CRY2 germline transformants were still in progress.
Thus far, germline transformants of the CIBN component have been generated, and work will continue to generate the CRY2 germline transformants.
Germline transformants of the CIBN component had been generated, while CRY2 germline transformants were still in progress.
Thus far, germline transformants of the CIBN component have been generated, and work will continue to generate the CRY2 germline transformants.
Blue light stimuli can selectively initiate protein-protein interactions in genetically encoded light-sensitive protein systems.
using blue light stimuli to selectively initiate protein-protein interactions
Blue light stimuli can selectively initiate protein-protein interactions in genetically encoded light-sensitive protein systems.
using blue light stimuli to selectively initiate protein-protein interactions
Blue light stimuli can selectively initiate protein-protein interactions in genetically encoded light-sensitive protein systems.
using blue light stimuli to selectively initiate protein-protein interactions
Blue light stimuli can selectively initiate protein-protein interactions in genetically encoded light-sensitive protein systems.
using blue light stimuli to selectively initiate protein-protein interactions
Blue light stimuli can selectively initiate protein-protein interactions in genetically encoded light-sensitive protein systems.
using blue light stimuli to selectively initiate protein-protein interactions
The CRY2/CIB module offers a genetically encoded mechanism to study protein roles in a tissue-specific manner during development.
the CRY2/CIB module, offers a powerful genetically encoded mechanism by which to study the role of proteins in a tissue-specific manner during various stages of development
The CRY2/CIB module offers a genetically encoded mechanism to study protein roles in a tissue-specific manner during development.
the CRY2/CIB module, offers a powerful genetically encoded mechanism by which to study the role of proteins in a tissue-specific manner during various stages of development
The CRY2/CIB module offers a genetically encoded mechanism to study protein roles in a tissue-specific manner during development.
the CRY2/CIB module, offers a powerful genetically encoded mechanism by which to study the role of proteins in a tissue-specific manner during various stages of development
The CRY2/CIB module offers a genetically encoded mechanism to study protein roles in a tissue-specific manner during development.
the CRY2/CIB module, offers a powerful genetically encoded mechanism by which to study the role of proteins in a tissue-specific manner during various stages of development
The CRY2/CIB module offers a genetically encoded mechanism to study protein roles in a tissue-specific manner during development.
the CRY2/CIB module, offers a powerful genetically encoded mechanism by which to study the role of proteins in a tissue-specific manner during various stages of development
Approval Evidence
1 source1 linked approval claimfirst-pass slug rho1-cry2-fusion-construct
working on cloning variants of the small G protein Rho to form a fusion protein with the CRY2 component
Source:
intended functionneutral
If Rho1 is successfully fused to CRY2, blue light could spatially and temporally control recruitment of CRY2 to membrane-anchored CIB and thereby affect downstream events.
If Drosophila Rho1 proteins are successfully adapted to CRY2 components, upon blue light stimulation the recruitment of CRY2 to a CIB component anchored in the membrane could be spatially and temporally controlled to affect subsequent downstream events.
Source:
Comparisons
Source-backed strengths
A key proposed strength is the use of blue light to drive reversible, spatially restricted recruitment through the established CRY2/CIB interaction framework. Another supported advantage is that the broader CRY2/CIB system was being adapted into Drosophila-specific vectors, which is relevant for organism-compatible implementation, but no performance data are provided for the Rho1 fusion itself.
Source:
Using cloning techniques to generate CRY2 and CIB constructs in Drosophila specific vectors, we have attempted to adapt the CRY2/CIB system to Drosophila.
Compared with blue light-activated PKC isozyme recruitment system
Rho1-CRY2 fusion construct and blue light-activated PKC isozyme recruitment system address a similar problem space because they share localization.
Shared frame: same top-level item type; shared target processes: localization; shared mechanisms: light-induced heterodimerization; same primary input modality: light
Rho1-CRY2 fusion construct and CRY2-talin/CIBN-CAAX optogenetic plasma membrane recruitment system address a similar problem space because they share localization.
Shared frame: same top-level item type; shared target processes: localization; shared mechanisms: light-induced heterodimerization; same primary input modality: light
Compared with iLID-antiGFP-nanobody
Rho1-CRY2 fusion construct and iLID-antiGFP-nanobody address a similar problem space because they share localization.
Shared frame: same top-level item type; shared target processes: localization; shared mechanisms: light-induced heterodimerization; same primary input modality: light
Ranked Citations
- 1.