Source 1primary paper2012Bioconjugate Chemistry
Toolkit/LOV-PvuII fusion enzyme
LOV-PvuII fusion enzyme
Also known as: light-controllable endonuclease, light-inducible chimeric endonucleases, LOV-PvuII variants
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The LOV-PvuII fusion enzyme is a genetically encoded light-controllable endonuclease created by fusing the Avena sativa phototropin1 LOV2 photosensory domain to the restriction enzyme PvuII. In analyzed variants, blue light modulated DNA cleavage activity relative to dark conditions, with the direction of regulation determined by the fusion interface.
Usefulness & Problems
Why this is useful
This tool provides optical control over restriction endonuclease activity using a genetically encoded protein fusion rather than an added chemical regulator. It is useful where reversible blue-light-dependent modulation of DNA cleavage is desired and where different fusion designs can bias activity toward either the dark or illuminated state.
Problem solved
It addresses the problem of making DNA cleavage by PvuII responsive to light. The reported chimeras convert blue-light input into altered nuclease activity, enabling conditional control of cleavage state through protein engineering at the fusion interface.
Problem links
Need conditional recombination or state switching
DerivedThe LOV-PvuII fusion enzyme is a genetically encoded light-controllable endonuclease created by fusing the Avena sativa phototropin1 LOV2 domain to the restriction enzyme PvuII. In analyzed variants, blue light modulated DNA cleavage activity relative to dark conditions, and the direction of regulation depended on the fusion interface.
Need precise spatiotemporal control with light input
DerivedThe LOV-PvuII fusion enzyme is a genetically encoded light-controllable endonuclease created by fusing the Avena sativa phototropin1 LOV2 domain to the restriction enzyme PvuII. In analyzed variants, blue light modulated DNA cleavage activity relative to dark conditions, and the direction of regulation depended on the fusion interface.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
light-dependent allosteric switchinglight-dependent allosteric switchingPhotocleavagereversible photoactivationreversible photoactivationTechniques
No technique tags yet.
Target processes
recombinationInput: 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: cleavageswitch architecture: multi componentswitch architecture: uncaging
The construct is implemented as a fusion between the Avena sativa phototropin1 LOV2 domain and the PvuII restriction enzyme. Blue-light illumination and dark conditions were the compared input states, and functional behavior depended on the specific fusion interface used; no additional practical details on expression, cofactors, or delivery are provided in the supplied evidence.
The available evidence is limited to a single cited study and a small set of analyzed variants. Quantitative performance is only reported as an approximately 3-fold light/dark activity difference, and the provided evidence does not describe validation in cells, genome engineering contexts, or recombination assays.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
DNA cleavage activity difference 3 fold
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
Approval Evidence
1 source4 linked approval claimsfirst-pass slug lov-pvuii-fusion-enzyme
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Source:
activity modulationsupports
Analyzed LOV-PvuII fusion variants showed a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions.
By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a 3-fold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark.
Source:
engineering resultsupports
Fusion of the Avena sativa phototropin1 LOV2 domain to PvuII generated a genetically encoded light-controllable endonuclease.
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Source:
mechanistic behaviorsupports
LOV-PvuII variants displayed bidirectional polarity in photoactivation depending on the fusion interface, with increased DNA cleavage activity occurring either in the dark state or in the blue-light photoexcited state.
Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photoactivation; i.e., increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photoexcitation state, when the LOV domain is partially unfolded.
Source:
reversibilitysupports
The light-dependent effect on LOV-PvuII fusion enzyme activity was fully reversible over multiple photocycles.
The effect is fully reversible over multiple photocycles.
Source:
Comparisons
Source-backed strengths
The reported fusion strategy successfully generated a light-controllable endonuclease from defined components, Avena sativa phototropin1 LOV2 and PvuII. Analyzed variants showed about a 3-fold difference in DNA cleavage activity between blue-light illumination and dark conditions, and the system supported bidirectional photoactivation polarity depending on fusion design.
Source:
Here, we have fused the light-sensitive LOV2 domain from Avena sativa phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease.
Compared with GFP-PHR-caspase8/Flag-CIB1N-caspase8
LOV-PvuII fusion enzyme and GFP-PHR-caspase8/Flag-CIB1N-caspase8 address a similar problem space because they share recombination.
Shared frame: same top-level item type; shared target processes: recombination; shared mechanisms: photocleavage; same primary input modality: light
Compared with PA-Cre 3.0
LOV-PvuII fusion enzyme and PA-Cre 3.0 address a similar problem space because they share recombination.
Shared frame: same top-level item type; shared target processes: recombination; shared mechanisms: photocleavage; same primary input modality: light
Compared with photocaged IPTG
LOV-PvuII fusion enzyme and photocaged IPTG address a similar problem space because they share recombination.
Shared frame: same top-level item type; shared target processes: recombination; shared mechanisms: photocleavage; same primary input modality: light
Ranked Citations
- 1.