Source 1primary paper2021Journal of the American Chemical Society
Toolkit/Blue Light Inducible SpyTag System
Blue Light Inducible SpyTag System
Also known as: BLISS
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
BLISS is a blue-light-inducible SpyTag system generated by inserting SpyTag into different positions of the AsLOV2 Jα-helix. In this design, blue light exposure enables conditional SpyTag reactivity with SpyCatcher, allowing light-gated coupling.
Usefulness & Problems
Why this is useful
BLISS provides optical control over SpyTag-SpyCatcher coupling, enabling spatially defined protein assembly. The source literature demonstrated this utility by photopatterning two fluorescent proteins and reported that the reaction could be quenched immediately by removing light.
Source:
We demonstrated the spatial aspect of this light control mechanism through photopatterning of two fluorescent proteins.
Problem solved
BLISS addresses the problem of making SpyTag-SpyCatcher association conditionally controllable by light rather than constitutively reactive. This supports experiments requiring spatial and temporal regulation of covalent protein coupling.
Source:
We demonstrated the spatial aspect of this light control mechanism through photopatterning of two fluorescent proteins.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
conditional spytag-spycatcher covalent associationconformational uncagingConformational Uncaginglight-induced jα-helix undockingTechniques
Computational DesignTarget processes
recombinationInput: Light
Implementation Constraints
BLISS was engineered by inserting SpyTag into different locations of the AsLOV2 Jα-helix. The supplied evidence supports blue light as the input modality, but does not provide construct sequences, illumination parameters, expression context, or delivery details.
The provided evidence does not report quantitative performance metrics such as dynamic range, kinetics, background reactivity in the dark, or performance across cell types. Validation in the supplied evidence is limited to the original study and a fluorescent protein photopatterning demonstration.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
The authors demonstrated spatial light control using BLISS by photopatterning two fluorescent proteins.
We demonstrated the spatial aspect of this light control mechanism through photopatterning of two fluorescent proteins.
The authors demonstrated spatial light control using BLISS by photopatterning two fluorescent proteins.
We demonstrated the spatial aspect of this light control mechanism through photopatterning of two fluorescent proteins.
The authors demonstrated spatial light control using BLISS by photopatterning two fluorescent proteins.
We demonstrated the spatial aspect of this light control mechanism through photopatterning of two fluorescent proteins.
The authors demonstrated spatial light control using BLISS by photopatterning two fluorescent proteins.
We demonstrated the spatial aspect of this light control mechanism through photopatterning of two fluorescent proteins.
The authors demonstrated spatial light control using BLISS by photopatterning two fluorescent proteins.
We demonstrated the spatial aspect of this light control mechanism through photopatterning of two fluorescent proteins.
The authors demonstrated spatial light control using BLISS by photopatterning two fluorescent proteins.
We demonstrated the spatial aspect of this light control mechanism through photopatterning of two fluorescent proteins.
The authors demonstrated spatial light control using BLISS by photopatterning two fluorescent proteins.
We demonstrated the spatial aspect of this light control mechanism through photopatterning of two fluorescent proteins.
The BLISS reaction could be instantaneously quenched by removing light.
Further, the reaction could be instantaneously quenched by removing light.
The BLISS reaction could be instantaneously quenched by removing light.
Further, the reaction could be instantaneously quenched by removing light.
The BLISS reaction could be instantaneously quenched by removing light.
Further, the reaction could be instantaneously quenched by removing light.
The BLISS reaction could be instantaneously quenched by removing light.
Further, the reaction could be instantaneously quenched by removing light.
The BLISS reaction could be instantaneously quenched by removing light.
Further, the reaction could be instantaneously quenched by removing light.
The BLISS reaction could be instantaneously quenched by removing light.
Further, the reaction could be instantaneously quenched by removing light.
The BLISS reaction could be instantaneously quenched by removing light.
Further, the reaction could be instantaneously quenched by removing light.
Insertion of SpyTag into different locations of the AsLOV2 Jα-helix created a blue-light-inducible SpyTag system called BLISS.
By inserting SpyTag into the different locations of the Jα-helix, we created a blue light inducible SpyTag system (BLISS).
Insertion of SpyTag into different locations of the AsLOV2 Jα-helix created a blue-light-inducible SpyTag system called BLISS.
By inserting SpyTag into the different locations of the Jα-helix, we created a blue light inducible SpyTag system (BLISS).
Insertion of SpyTag into different locations of the AsLOV2 Jα-helix created a blue-light-inducible SpyTag system called BLISS.
By inserting SpyTag into the different locations of the Jα-helix, we created a blue light inducible SpyTag system (BLISS).
Insertion of SpyTag into different locations of the AsLOV2 Jα-helix created a blue-light-inducible SpyTag system called BLISS.
By inserting SpyTag into the different locations of the Jα-helix, we created a blue light inducible SpyTag system (BLISS).
Insertion of SpyTag into different locations of the AsLOV2 Jα-helix created a blue-light-inducible SpyTag system called BLISS.
By inserting SpyTag into the different locations of the Jα-helix, we created a blue light inducible SpyTag system (BLISS).
Insertion of SpyTag into different locations of the AsLOV2 Jα-helix created a blue-light-inducible SpyTag system called BLISS.
By inserting SpyTag into the different locations of the Jα-helix, we created a blue light inducible SpyTag system (BLISS).
Insertion of SpyTag into different locations of the AsLOV2 Jα-helix created a blue-light-inducible SpyTag system called BLISS.
By inserting SpyTag into the different locations of the Jα-helix, we created a blue light inducible SpyTag system (BLISS).
In BLISS, SpyTag is blocked from reacting with SpyCatcher in the dark, and blue-light irradiation exposes SpyTag through AsLOV2 Jα-helix undocking.
In this design, the SpyTag is blocked from reacting with the SpyCatcher in the dark, but upon irradiation with blue light, the Jα-helix of the AsLOV2 undocks to expose the SpyTag.
In BLISS, SpyTag is blocked from reacting with SpyCatcher in the dark, and blue-light irradiation exposes SpyTag through AsLOV2 Jα-helix undocking.
In this design, the SpyTag is blocked from reacting with the SpyCatcher in the dark, but upon irradiation with blue light, the Jα-helix of the AsLOV2 undocks to expose the SpyTag.
In BLISS, SpyTag is blocked from reacting with SpyCatcher in the dark, and blue-light irradiation exposes SpyTag through AsLOV2 Jα-helix undocking.
In this design, the SpyTag is blocked from reacting with the SpyCatcher in the dark, but upon irradiation with blue light, the Jα-helix of the AsLOV2 undocks to expose the SpyTag.
In BLISS, SpyTag is blocked from reacting with SpyCatcher in the dark, and blue-light irradiation exposes SpyTag through AsLOV2 Jα-helix undocking.
In this design, the SpyTag is blocked from reacting with the SpyCatcher in the dark, but upon irradiation with blue light, the Jα-helix of the AsLOV2 undocks to expose the SpyTag.
In BLISS, SpyTag is blocked from reacting with SpyCatcher in the dark, and blue-light irradiation exposes SpyTag through AsLOV2 Jα-helix undocking.
In this design, the SpyTag is blocked from reacting with the SpyCatcher in the dark, but upon irradiation with blue light, the Jα-helix of the AsLOV2 undocks to expose the SpyTag.
In BLISS, SpyTag is blocked from reacting with SpyCatcher in the dark, and blue-light irradiation exposes SpyTag through AsLOV2 Jα-helix undocking.
In this design, the SpyTag is blocked from reacting with the SpyCatcher in the dark, but upon irradiation with blue light, the Jα-helix of the AsLOV2 undocks to expose the SpyTag.
In BLISS, SpyTag is blocked from reacting with SpyCatcher in the dark, and blue-light irradiation exposes SpyTag through AsLOV2 Jα-helix undocking.
In this design, the SpyTag is blocked from reacting with the SpyCatcher in the dark, but upon irradiation with blue light, the Jα-helix of the AsLOV2 undocks to expose the SpyTag.
The authors found three BLISS variants with dynamic ranges greater than 15 and activity in different concentration ranges.
We found three variants with dynamic ranges over 15, which were active within different concentration ranges.
dynamic range 15
The authors found three BLISS variants with dynamic ranges greater than 15 and activity in different concentration ranges.
We found three variants with dynamic ranges over 15, which were active within different concentration ranges.
dynamic range 15
The authors found three BLISS variants with dynamic ranges greater than 15 and activity in different concentration ranges.
We found three variants with dynamic ranges over 15, which were active within different concentration ranges.
dynamic range 15
The authors found three BLISS variants with dynamic ranges greater than 15 and activity in different concentration ranges.
We found three variants with dynamic ranges over 15, which were active within different concentration ranges.
dynamic range 15
The authors found three BLISS variants with dynamic ranges greater than 15 and activity in different concentration ranges.
We found three variants with dynamic ranges over 15, which were active within different concentration ranges.
dynamic range 15
The authors found three BLISS variants with dynamic ranges greater than 15 and activity in different concentration ranges.
We found three variants with dynamic ranges over 15, which were active within different concentration ranges.
dynamic range 15
The authors found three BLISS variants with dynamic ranges greater than 15 and activity in different concentration ranges.
We found three variants with dynamic ranges over 15, which were active within different concentration ranges.
dynamic range 15
BLISS activity could be tuned using SpyCatcher variants with different reaction kinetics.
These could be tuned using SpyCatcher variants with different reaction kinetics.
BLISS activity could be tuned using SpyCatcher variants with different reaction kinetics.
These could be tuned using SpyCatcher variants with different reaction kinetics.
BLISS activity could be tuned using SpyCatcher variants with different reaction kinetics.
These could be tuned using SpyCatcher variants with different reaction kinetics.
BLISS activity could be tuned using SpyCatcher variants with different reaction kinetics.
These could be tuned using SpyCatcher variants with different reaction kinetics.
BLISS activity could be tuned using SpyCatcher variants with different reaction kinetics.
These could be tuned using SpyCatcher variants with different reaction kinetics.
BLISS activity could be tuned using SpyCatcher variants with different reaction kinetics.
These could be tuned using SpyCatcher variants with different reaction kinetics.
BLISS activity could be tuned using SpyCatcher variants with different reaction kinetics.
These could be tuned using SpyCatcher variants with different reaction kinetics.
Approval Evidence
1 source6 linked approval claimsfirst-pass slug blue-light-inducible-spytag-system
By inserting SpyTag into the different locations of the Jα-helix, we created a blue light inducible SpyTag system (BLISS).
Source:
application demosupports
The authors demonstrated spatial light control using BLISS by photopatterning two fluorescent proteins.
We demonstrated the spatial aspect of this light control mechanism through photopatterning of two fluorescent proteins.
Source:
control propertysupports
The BLISS reaction could be instantaneously quenched by removing light.
Further, the reaction could be instantaneously quenched by removing light.
Source:
engineering resultsupports
Insertion of SpyTag into different locations of the AsLOV2 Jα-helix created a blue-light-inducible SpyTag system called BLISS.
By inserting SpyTag into the different locations of the Jα-helix, we created a blue light inducible SpyTag system (BLISS).
Source:
mechanismsupports
In BLISS, SpyTag is blocked from reacting with SpyCatcher in the dark, and blue-light irradiation exposes SpyTag through AsLOV2 Jα-helix undocking.
In this design, the SpyTag is blocked from reacting with the SpyCatcher in the dark, but upon irradiation with blue light, the Jα-helix of the AsLOV2 undocks to expose the SpyTag.
Source:
performancesupports
The authors found three BLISS variants with dynamic ranges greater than 15 and activity in different concentration ranges.
We found three variants with dynamic ranges over 15, which were active within different concentration ranges.
Source:
tunabilitysupports
BLISS activity could be tuned using SpyCatcher variants with different reaction kinetics.
These could be tuned using SpyCatcher variants with different reaction kinetics.
Source:
Comparisons
Source-backed strengths
The reported system is blue-light inducible and supports spatial light control, as shown by photopatterning of two fluorescent proteins. The reaction was also reported to be instantaneously quenched upon light removal, indicating reversible optical control over the inducible state.
Source:
By inserting SpyTag into the different locations of the Jα-helix, we created a blue light inducible SpyTag system (BLISS).
Source:
We found three variants with dynamic ranges over 15, which were active within different concentration ranges.
Ranked Citations
- 1.