Source 1primary paper2019
Toolkit/LOV2 blue light sensory domain
LOV2 blue light sensory domain
Also known as: LOV2
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
LOV2 is a blue-light sensory domain from Avena sativa that was fused into the anti-CRISPR protein AcrIIC3 to create light-switchable inhibitors of Neisseria meningitidis Cas9. In this engineered context, AcrIIC3-LOV2 hybrids inhibited Nme Cas9 in the dark and permitted genome editing under blue light in mammalian cells.
Usefulness & Problems
Why this is useful
This domain is useful as an optogenetic module for converting a constitutive anti-CRISPR protein into a light-responsive regulator of Cas9 activity. The reported application enables temporal control of type II-C CRISPR effector function using blue light.
Source:
Here, we report the first optogenetic tool to control Nme Cas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein.
Problem solved
The engineered LOV2 insertion addressed the problem of externally controlling Nme Cas9 genome editing with light rather than constitutive inhibition. The study specifically demonstrated a route to design optogenetic anti-CRISPR proteins for reversible activity gating.
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
No target processes tagged yet.
Input: Light
Implementation Constraints
The validated implementation involved domain fusion of LOV2 into AcrIIC3 to generate light-switchable anti-CRISPR hybrids. Blue light was the input modality, and the reported output was dark-state inhibition with light-permissive genome editing in mammalian cells; no additional construct, cofactor, or delivery details are provided in the supplied evidence.
The supplied evidence is limited to a single 2019 study and one engineered use case involving AcrIIC3 and Nme Cas9 in mammalian cells. No independent replication, quantitative performance metrics, or broader validation across other proteins, organisms, or illumination regimes are provided here.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Two AcrIIC3-LOV2 hybrids blocked Nme Cas9 activity in the dark and permitted genome editing upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids from our collection potently blocked Nme Cas9 activity in the dark, while permitting robust genome editing at various endogenous loci upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids blocked Nme Cas9 activity in the dark and permitted genome editing upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids from our collection potently blocked Nme Cas9 activity in the dark, while permitting robust genome editing at various endogenous loci upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids blocked Nme Cas9 activity in the dark and permitted genome editing upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids from our collection potently blocked Nme Cas9 activity in the dark, while permitting robust genome editing at various endogenous loci upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids blocked Nme Cas9 activity in the dark and permitted genome editing upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids from our collection potently blocked Nme Cas9 activity in the dark, while permitting robust genome editing at various endogenous loci upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids blocked Nme Cas9 activity in the dark and permitted genome editing upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids from our collection potently blocked Nme Cas9 activity in the dark, while permitting robust genome editing at various endogenous loci upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids blocked Nme Cas9 activity in the dark and permitted genome editing upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids from our collection potently blocked Nme Cas9 activity in the dark, while permitting robust genome editing at various endogenous loci upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids blocked Nme Cas9 activity in the dark and permitted genome editing upon blue light irradiation.
Two AcrIIC3-LOV2 hybrids from our collection potently blocked Nme Cas9 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.
Structural analysis placed the LOV2 domain in close proximity to the Cas9 binding surface within the hybrids.
Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface.
Structural analysis placed the LOV2 domain in close proximity to the Cas9 binding surface within the hybrids.
Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface.
Structural analysis placed the LOV2 domain in close proximity to the Cas9 binding surface within the hybrids.
Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface.
Structural analysis placed the LOV2 domain in close proximity to the Cas9 binding surface within the hybrids.
Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface.
Structural analysis placed the LOV2 domain in close proximity to the Cas9 binding surface within the hybrids.
Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface.
Structural analysis placed the LOV2 domain in close proximity to the Cas9 binding surface within the hybrids.
Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface.
Structural analysis placed the LOV2 domain in close proximity to the Cas9 binding surface within the hybrids.
Structural analysis revealed that, within these hybrids, the LOV2 domain is located in striking proximity to the Cas9 binding surface.
This work reports an optogenetic tool that controls Nme Cas9 activity in mammalian cells using an engineered light-dependent anti-CRISPR protein.
Here, we report the first optogenetic tool to control Nme Cas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein.
This work reports an optogenetic tool that controls Nme Cas9 activity in mammalian cells using an engineered light-dependent anti-CRISPR protein.
Here, we report the first optogenetic tool to control Nme Cas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein.
This work reports an optogenetic tool that controls Nme Cas9 activity in mammalian cells using an engineered light-dependent anti-CRISPR protein.
Here, we report the first optogenetic tool to control Nme Cas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein.
This work reports an optogenetic tool that controls Nme Cas9 activity in mammalian cells using an engineered light-dependent anti-CRISPR protein.
Here, we report the first optogenetic tool to control Nme Cas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein.
This work reports an optogenetic tool that controls Nme Cas9 activity in mammalian cells using an engineered light-dependent anti-CRISPR protein.
Here, we report the first optogenetic tool to control Nme Cas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein.
This work reports an optogenetic tool that controls Nme Cas9 activity in mammalian cells using an engineered light-dependent anti-CRISPR protein.
Here, we report the first optogenetic tool to control Nme Cas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein.
This work reports an optogenetic tool that controls Nme Cas9 activity in mammalian cells using an engineered light-dependent anti-CRISPR protein.
Here, we report the first optogenetic tool to control Nme Cas9 activity in mammalian cells via an engineered, light-dependent anti-CRISPR (Acr) protein.
Approval Evidence
1 source1 linked approval claimfirst-pass slug lov2-blue-light-sensory-domain
the LOV2 blue light sensory domain from Avena sativa
Source:
structural observationsupports
Structural analysis placed the LOV2 domain in close proximity to the Cas9 binding surface within the hybrids.
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
Two AcrIIC3-LOV2 hybrids were reported to block Nme Cas9 activity in the dark while allowing genome editing upon blue-light irradiation. Structural analysis further showed that the inserted LOV2 domain was positioned close to the Cas9-binding surface in the hybrids, supporting the functional design logic.
Ranked Citations
- 1.