Source 1primary paper2020Nucleic Acids Research
Toolkit/LOV2 blue light sensory domain from Avena sativa
LOV2 blue light sensory domain from Avena sativa
Also known as: LOV2
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The Avena sativa LOV2 blue light sensory domain is a photosensory protein module used in engineered AcrIIC3-LOV2 fusion proteins to confer blue-light control over Neisseria meningitidis Cas9 activity. In this reported context, LOV2 enabled dark-state inhibition and light-permissive genome editing in mammalian cells.
Usefulness & Problems
Why this is useful
This domain is useful as an optogenetic module for making anti-CRISPR function responsive to blue light. The cited study indicates that LOV2 can support reversible activity gating of a type II-C CRISPR effector through fusion to AcrIIC3.
Problem solved
It addresses the problem of controlling NmeCas9 genome editing with light rather than constitutive inhibition. Specifically, the engineered hybrids allowed potent blocking in the dark while permitting robust editing under blue light irradiation.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level protein part used inside a larger architecture that realizes a mechanism.
Techniques
Structural CharacterizationTarget processes
recombinationInput: Light
Implementation Constraints
The reported implementation used the Avena sativa LOV2 blue light sensory domain as a fusion partner in AcrIIC3-LOV2 hybrid proteins. The validated application involved blue light irradiation, anti-CRISPR-mediated control of NmeCas9, and testing in mammalian cells.
The supplied evidence only supports use of LOV2 in AcrIIC3-LOV2 hybrids regulating NmeCas9, so broader portability to other proteins, organisms, or editing systems is not established here. Quantitative performance metrics, kinetics, and independent replication are not provided in the supplied evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Two AcrIIC3-LOV2 hybrids potently blocked NmeCas9 activity in the dark while permitting robust genome editing upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids from our collection potently blocked NmeCas9 activity in the dark, while permitting robust genome editing at various endogenous loci upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids potently blocked NmeCas9 activity in the dark while permitting robust genome editing upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids from our collection potently blocked NmeCas9 activity in the dark, while permitting robust genome editing at various endogenous loci upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids potently blocked NmeCas9 activity in the dark while permitting robust genome editing upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids from our collection potently blocked NmeCas9 activity in the dark, while permitting robust genome editing at various endogenous loci upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids potently blocked NmeCas9 activity in the dark while permitting robust genome editing upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids from our collection potently blocked NmeCas9 activity in the dark, while permitting robust genome editing at various endogenous loci upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids potently blocked NmeCas9 activity in the dark while permitting robust genome editing upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids from our collection potently blocked NmeCas9 activity in the dark, while permitting robust genome editing at various endogenous loci upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids potently blocked NmeCas9 activity in the dark while permitting robust genome editing upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids from our collection potently blocked NmeCas9 activity in the dark, while permitting robust genome editing at various endogenous loci upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids potently blocked NmeCas9 activity in the dark while permitting robust genome editing upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids from our collection potently blocked NmeCas9 activity in the dark, while permitting robust genome editing at various endogenous loci upon blue light irradiation.
The work demonstrates optogenetic regulation of a type II-C CRISPR effector and suggests a route for designing optogenetic anti-CRISPR proteins.
Together, our work demonstrates optogenetic regulation of a type II-C CRISPR effector and might suggest a new route for the design of optogenetic Acrs.
The work demonstrates optogenetic regulation of a type II-C CRISPR effector and suggests a route for designing optogenetic anti-CRISPR proteins.
Together, our work demonstrates optogenetic regulation of a type II-C CRISPR effector and might suggest a new route for the design of optogenetic Acrs.
The work demonstrates optogenetic regulation of a type II-C CRISPR effector and suggests a route for designing optogenetic anti-CRISPR proteins.
Together, our work demonstrates optogenetic regulation of a type II-C CRISPR effector and might suggest a new route for the design of optogenetic Acrs.
The work demonstrates optogenetic regulation of a type II-C CRISPR effector and suggests a route for designing optogenetic anti-CRISPR proteins.
Together, our work demonstrates optogenetic regulation of a type II-C CRISPR effector and might suggest a new route for the design of optogenetic Acrs.
The work demonstrates optogenetic regulation of a type II-C CRISPR effector and suggests a route for designing optogenetic anti-CRISPR proteins.
Together, our work demonstrates optogenetic regulation of a type II-C CRISPR effector and might suggest a new route for the design of optogenetic Acrs.
The work demonstrates optogenetic regulation of a type II-C CRISPR effector and suggests a route for designing optogenetic anti-CRISPR proteins.
Together, our work demonstrates optogenetic regulation of a type II-C CRISPR effector and might suggest a new route for the design of optogenetic Acrs.
The work demonstrates optogenetic regulation of a type II-C CRISPR effector and suggests a route for designing optogenetic anti-CRISPR proteins.
Together, our work demonstrates optogenetic regulation of a type II-C CRISPR effector and might suggest a new route for the design of optogenetic Acrs.
The authors created hybrids between AcrIIC3 and the Avena sativa LOV2 blue light sensory domain.
we created hybrids between the NmeCas9 inhibitor AcrIIC3 and the LOV2 blue light sensory domain from Avena sativa
The authors created hybrids between AcrIIC3 and the Avena sativa LOV2 blue light sensory domain.
we created hybrids between the NmeCas9 inhibitor AcrIIC3 and the LOV2 blue light sensory domain from Avena sativa
The authors created hybrids between AcrIIC3 and the Avena sativa LOV2 blue light sensory domain.
we created hybrids between the NmeCas9 inhibitor AcrIIC3 and the LOV2 blue light sensory domain from Avena sativa
The authors created hybrids between AcrIIC3 and the Avena sativa LOV2 blue light sensory domain.
we created hybrids between the NmeCas9 inhibitor AcrIIC3 and the LOV2 blue light sensory domain from Avena sativa
The authors created hybrids between AcrIIC3 and the Avena sativa LOV2 blue light sensory domain.
we created hybrids between the NmeCas9 inhibitor AcrIIC3 and the LOV2 blue light sensory domain from Avena sativa
The authors created hybrids between AcrIIC3 and the Avena sativa LOV2 blue light sensory domain.
we created hybrids between the NmeCas9 inhibitor AcrIIC3 and the LOV2 blue light sensory domain from Avena sativa
The authors created hybrids between AcrIIC3 and the Avena sativa LOV2 blue light sensory domain.
we created hybrids between the NmeCas9 inhibitor AcrIIC3 and the LOV2 blue light sensory domain from Avena sativa
This paper reports the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered light-dependent anti-CRISPR protein.
Here, we report the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein.
This paper reports the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered light-dependent anti-CRISPR protein.
Here, we report the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein.
This paper reports the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered light-dependent anti-CRISPR protein.
Here, we report the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein.
This paper reports the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered light-dependent anti-CRISPR protein.
Here, we report the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein.
This paper reports the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered light-dependent anti-CRISPR protein.
Here, we report the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein.
This paper reports the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered light-dependent anti-CRISPR protein.
Here, we report the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein.
This paper reports the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered light-dependent anti-CRISPR protein.
Here, we report the first optogenetic tool to control NmeCas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein.
Structural analysis indicated that the LOV2 domain in the hybrids is located close to the Cas9 binding surface.
Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface.
Structural analysis indicated that the LOV2 domain in the hybrids is located close to the Cas9 binding surface.
Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface.
Structural analysis indicated that the LOV2 domain in the hybrids is located close to the Cas9 binding surface.
Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface.
Structural analysis indicated that the LOV2 domain in the hybrids is located close to the Cas9 binding surface.
Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface.
Structural analysis indicated that the LOV2 domain in the hybrids is located close to the Cas9 binding surface.
Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface.
Structural analysis indicated that the LOV2 domain in the hybrids is located close to the Cas9 binding surface.
Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface.
Structural analysis indicated that the LOV2 domain in the hybrids is located close to the Cas9 binding surface.
Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface.
Approval Evidence
1 source2 linked approval claimsfirst-pass slug lov2-blue-light-sensory-domain-from-avena-sativa
the LOV2 blue light sensory domain from Avena sativa
Source:
engineering designsupports
The authors created hybrids between AcrIIC3 and the Avena sativa LOV2 blue light sensory domain.
we created hybrids between the NmeCas9 inhibitor AcrIIC3 and the LOV2 blue light sensory domain from Avena sativa
Source:
structural observationsupports
Structural analysis indicated that the LOV2 domain in the hybrids is located close to the Cas9 binding surface.
Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface.
Source:
Comparisons
Source-backed strengths
In the cited study, two AcrIIC3-LOV2 hybrids potently blocked NmeCas9 activity in the dark and permitted robust genome editing upon blue light exposure. The work also demonstrated optogenetic regulation of a type II-C CRISPR effector in mammalian cells.
Source:
we created hybrids between the NmeCas9 inhibitor AcrIIC3 and the LOV2 blue light sensory domain from Avena sativa
Ranked Citations
- 1.