Source 1primary paper2017Proceedings of the National Academy of Sciences
Toolkit/PCB synthesis expression vector
PCB synthesis expression vector
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The PCB synthesis expression vector is a mammalian multigene construct that coexpresses heme oxygenase 1, phycocyanobilin:ferredoxin oxidoreductase, ferredoxin, and ferredoxin-NADP+ reductase to drive phycocyanobilin synthesis in mitochondria. It is used to supply the chromophore required for PhyB-PIF optogenetic systems without external PCB supplementation.
Usefulness & Problems
Why this is useful
This construct is useful because it enables intracellular production of phycocyanobilin in mammalian cells, removing the need for exogenous chromophore addition when using PhyB-PIF optogenetic regulation. The cited application is optogenetic control of intracellular signaling.
Source:
The PCB synthesis and PhyB-PIF systems allowed us to optogenetically regulate intracellular signaling without any external supply of chromophores.
Source:
Here, we report an expression vector that coexpresses HO1 and PcyA with Ferredoxin and Ferredoxin-NADP+ reductase for the efficient synthesis of PCB in the mitochondria of mammalian cells.
Problem solved
A key limitation of PhyB-PIF optogenetics in mammalian cells is the lack of endogenous phycocyanobilin production. This vector addresses that problem by reconstituting the enzymatic pathway for PCB synthesis in mitochondria.
Source:
The PCB synthesis and PhyB-PIF systems allowed us to optogenetically regulate intracellular signaling without any external supply of chromophores.
Source:
Here, we report an expression vector that coexpresses HO1 and PcyA with Ferredoxin and Ferredoxin-NADP+ reductase for the efficient synthesis of PCB in the mitochondria of mammalian cells.
Problem links
Need conditional control of signaling activity
DerivedThe PCB synthesis expression vector is a mammalian multigene construct that coexpresses heme oxygenase 1, phycocyanobilin:ferredoxin oxidoreductase, ferredoxin, and ferredoxin-NADP+ reductase to drive phycocyanobilin synthesis in mitochondria. It is used to supply the chromophore required for PhyB-PIF optogenetic systems without external PCB supplementation.
Need conditional protein clearance
DerivedThe PCB synthesis expression vector is a mammalian multigene construct that coexpresses heme oxygenase 1, phycocyanobilin:ferredoxin oxidoreductase, ferredoxin, and ferredoxin-NADP+ reductase to drive phycocyanobilin synthesis in mitochondria. It is used to supply the chromophore required for PhyB-PIF optogenetic systems without external PCB supplementation.
Need conditional recombination or state switching
DerivedThe PCB synthesis expression vector is a mammalian multigene construct that coexpresses heme oxygenase 1, phycocyanobilin:ferredoxin oxidoreductase, ferredoxin, and ferredoxin-NADP+ reductase to drive phycocyanobilin synthesis in mitochondria. It is used to supply the chromophore required for PhyB-PIF optogenetic systems without external PCB supplementation.
Need precise spatiotemporal control with light input
DerivedThe PCB synthesis expression vector is a mammalian multigene construct that coexpresses heme oxygenase 1, phycocyanobilin:ferredoxin oxidoreductase, ferredoxin, and ferredoxin-NADP+ reductase to drive phycocyanobilin synthesis in mitochondria. It is used to supply the chromophore required for PhyB-PIF optogenetic systems without external PCB supplementation.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Techniques
No technique tags yet.
Target processes
degradationrecombinationsignalingInput: Light
Implementation Constraints
cofactor dependency: requires exogenous cofactorencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: spectral hardware requirementoperating role: regulator
The construct pattern involves multigene coexpression of HO1, PcyA, ferredoxin, and ferredoxin-NADP+ reductase in mammalian cells, with mitochondrial localization specified in the source text. The evidence also indicates that intracellular PCB levels can be increased by depletion of biliverdin reductase A.
The supplied evidence is limited to a single 2017 study and does not provide detailed quantitative performance metrics, cell-type scope, or long-term stability data. Independent replication is not documented in the provided evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
The PCB synthesis system together with the PhyB-PIF system enables optogenetic regulation of intracellular signaling without external chromophore supply.
The PCB synthesis and PhyB-PIF systems allowed us to optogenetically regulate intracellular signaling without any external supply of chromophores.
The PCB synthesis system together with the PhyB-PIF system enables optogenetic regulation of intracellular signaling without external chromophore supply.
The PCB synthesis and PhyB-PIF systems allowed us to optogenetically regulate intracellular signaling without any external supply of chromophores.
The PCB synthesis system together with the PhyB-PIF system enables optogenetic regulation of intracellular signaling without external chromophore supply.
The PCB synthesis and PhyB-PIF systems allowed us to optogenetically regulate intracellular signaling without any external supply of chromophores.
The PCB synthesis system together with the PhyB-PIF system enables optogenetic regulation of intracellular signaling without external chromophore supply.
The PCB synthesis and PhyB-PIF systems allowed us to optogenetically regulate intracellular signaling without any external supply of chromophores.
The PCB synthesis system together with the PhyB-PIF system enables optogenetic regulation of intracellular signaling without external chromophore supply.
The PCB synthesis and PhyB-PIF systems allowed us to optogenetically regulate intracellular signaling without any external supply of chromophores.
The PCB synthesis system together with the PhyB-PIF system enables optogenetic regulation of intracellular signaling without external chromophore supply.
The PCB synthesis and PhyB-PIF systems allowed us to optogenetically regulate intracellular signaling without any external supply of chromophores.
The PCB synthesis system together with the PhyB-PIF system enables optogenetic regulation of intracellular signaling without external chromophore supply.
The PCB synthesis and PhyB-PIF systems allowed us to optogenetically regulate intracellular signaling without any external supply of chromophores.
Depletion of biliverdin reductase A increases intracellular phycocyanobilin concentration.
An even higher intracellular PCB concentration was achieved by the depletion of biliverdin reductase A, which degrades PCB.
Depletion of biliverdin reductase A increases intracellular phycocyanobilin concentration.
An even higher intracellular PCB concentration was achieved by the depletion of biliverdin reductase A, which degrades PCB.
Depletion of biliverdin reductase A increases intracellular phycocyanobilin concentration.
An even higher intracellular PCB concentration was achieved by the depletion of biliverdin reductase A, which degrades PCB.
Depletion of biliverdin reductase A increases intracellular phycocyanobilin concentration.
An even higher intracellular PCB concentration was achieved by the depletion of biliverdin reductase A, which degrades PCB.
Depletion of biliverdin reductase A increases intracellular phycocyanobilin concentration.
An even higher intracellular PCB concentration was achieved by the depletion of biliverdin reductase A, which degrades PCB.
Depletion of biliverdin reductase A increases intracellular phycocyanobilin concentration.
An even higher intracellular PCB concentration was achieved by the depletion of biliverdin reductase A, which degrades PCB.
Depletion of biliverdin reductase A increases intracellular phycocyanobilin concentration.
An even higher intracellular PCB concentration was achieved by the depletion of biliverdin reductase A, which degrades PCB.
This work provides a practical method for a fully genetically encoded PhyB-PIF system.
Thus, we have provided a practical method for developing a fully genetically encoded PhyB-PIF system, which paves the way for its application to a living animal.
This work provides a practical method for a fully genetically encoded PhyB-PIF system.
Thus, we have provided a practical method for developing a fully genetically encoded PhyB-PIF system, which paves the way for its application to a living animal.
This work provides a practical method for a fully genetically encoded PhyB-PIF system.
Thus, we have provided a practical method for developing a fully genetically encoded PhyB-PIF system, which paves the way for its application to a living animal.
This work provides a practical method for a fully genetically encoded PhyB-PIF system.
Thus, we have provided a practical method for developing a fully genetically encoded PhyB-PIF system, which paves the way for its application to a living animal.
This work provides a practical method for a fully genetically encoded PhyB-PIF system.
Thus, we have provided a practical method for developing a fully genetically encoded PhyB-PIF system, which paves the way for its application to a living animal.
This work provides a practical method for a fully genetically encoded PhyB-PIF system.
Thus, we have provided a practical method for developing a fully genetically encoded PhyB-PIF system, which paves the way for its application to a living animal.
This work provides a practical method for a fully genetically encoded PhyB-PIF system.
Thus, we have provided a practical method for developing a fully genetically encoded PhyB-PIF system, which paves the way for its application to a living animal.
An expression vector coexpressing HO1, PcyA, ferredoxin, and ferredoxin-NADP+ reductase enables efficient synthesis of phycocyanobilin in mammalian cell mitochondria.
Here, we report an expression vector that coexpresses HO1 and PcyA with Ferredoxin and Ferredoxin-NADP+ reductase for the efficient synthesis of PCB in the mitochondria of mammalian cells.
An expression vector coexpressing HO1, PcyA, ferredoxin, and ferredoxin-NADP+ reductase enables efficient synthesis of phycocyanobilin in mammalian cell mitochondria.
Here, we report an expression vector that coexpresses HO1 and PcyA with Ferredoxin and Ferredoxin-NADP+ reductase for the efficient synthesis of PCB in the mitochondria of mammalian cells.
An expression vector coexpressing HO1, PcyA, ferredoxin, and ferredoxin-NADP+ reductase enables efficient synthesis of phycocyanobilin in mammalian cell mitochondria.
Here, we report an expression vector that coexpresses HO1 and PcyA with Ferredoxin and Ferredoxin-NADP+ reductase for the efficient synthesis of PCB in the mitochondria of mammalian cells.
An expression vector coexpressing HO1, PcyA, ferredoxin, and ferredoxin-NADP+ reductase enables efficient synthesis of phycocyanobilin in mammalian cell mitochondria.
Here, we report an expression vector that coexpresses HO1 and PcyA with Ferredoxin and Ferredoxin-NADP+ reductase for the efficient synthesis of PCB in the mitochondria of mammalian cells.
An expression vector coexpressing HO1, PcyA, ferredoxin, and ferredoxin-NADP+ reductase enables efficient synthesis of phycocyanobilin in mammalian cell mitochondria.
Here, we report an expression vector that coexpresses HO1 and PcyA with Ferredoxin and Ferredoxin-NADP+ reductase for the efficient synthesis of PCB in the mitochondria of mammalian cells.
An expression vector coexpressing HO1, PcyA, ferredoxin, and ferredoxin-NADP+ reductase enables efficient synthesis of phycocyanobilin in mammalian cell mitochondria.
Here, we report an expression vector that coexpresses HO1 and PcyA with Ferredoxin and Ferredoxin-NADP+ reductase for the efficient synthesis of PCB in the mitochondria of mammalian cells.
An expression vector coexpressing HO1, PcyA, ferredoxin, and ferredoxin-NADP+ reductase enables efficient synthesis of phycocyanobilin in mammalian cell mitochondria.
Here, we report an expression vector that coexpresses HO1 and PcyA with Ferredoxin and Ferredoxin-NADP+ reductase for the efficient synthesis of PCB in the mitochondria of mammalian cells.
Approval Evidence
1 source4 linked approval claimsfirst-pass slug pcb-synthesis-expression-vector
Here, we report an expression vector that coexpresses HO1 and PcyA with Ferredoxin and Ferredoxin-NADP+ reductase for the efficient synthesis of PCB in the mitochondria of mammalian cells.
Source:
applicationsupports
The PCB synthesis system together with the PhyB-PIF system enables optogenetic regulation of intracellular signaling without external chromophore supply.
The PCB synthesis and PhyB-PIF systems allowed us to optogenetically regulate intracellular signaling without any external supply of chromophores.
Source:
performance modulationsupports
Depletion of biliverdin reductase A increases intracellular phycocyanobilin concentration.
An even higher intracellular PCB concentration was achieved by the depletion of biliverdin reductase A, which degrades PCB.
Source:
practical methodsupports
This work provides a practical method for a fully genetically encoded PhyB-PIF system.
Thus, we have provided a practical method for developing a fully genetically encoded PhyB-PIF system, which paves the way for its application to a living animal.
Source:
tool developmentsupports
An expression vector coexpressing HO1, PcyA, ferredoxin, and ferredoxin-NADP+ reductase enables efficient synthesis of phycocyanobilin in mammalian cell mitochondria.
Here, we report an expression vector that coexpresses HO1 and PcyA with Ferredoxin and Ferredoxin-NADP+ reductase for the efficient synthesis of PCB in the mitochondria of mammalian cells.
Source:
Comparisons
Source-backed strengths
The reported design coexpresses all four required components—HO1, PcyA, ferredoxin, and ferredoxin-NADP+ reductase—in a single mitochondrial-targeted expression system for efficient PCB synthesis. The source literature further states that this PCB synthesis system supports optogenetic regulation of intracellular signaling without external chromophore supply, and that biliverdin reductase A depletion increases intracellular PCB concentration.
Source:
An even higher intracellular PCB concentration was achieved by the depletion of biliverdin reductase A, which degrades PCB.
Compared with constitutive SsDHN overexpression in transgenic tobacco
PCB synthesis expression vector and constitutive SsDHN overexpression in transgenic tobacco address a similar problem space because they share degradation, recombination.
Shared frame: same top-level item type; shared target processes: degradation, recombination; shared mechanisms: degradation; same primary input modality: light
Relative tradeoffs: looks easier to implement in practice; may avoid an exogenous cofactor requirement.
PCB synthesis expression vector and Jalpha helix point mutations disrupting LOV-domain interaction address a similar problem space because they share degradation, signaling.
Shared frame: same top-level item type; shared target processes: degradation, signaling; shared mechanisms: degradation; same primary input modality: light
Relative tradeoffs: looks easier to implement in practice; may avoid an exogenous cofactor requirement.
Compared with lyso-ArchT
PCB synthesis expression vector and lyso-ArchT address a similar problem space because they share degradation.
Shared frame: same top-level item type; shared target processes: degradation; shared mechanisms: degradation; same primary input modality: light
Relative tradeoffs: looks easier to implement in practice; may avoid an exogenous cofactor requirement.
Ranked Citations
- 1.