Source 1primary paper2020
Toolkit/drug inducible lentiviral and transposon vectors
drug inducible lentiviral and transposon vectors
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Drug-inducible lentiviral and transposon vectors were used to deliver the PhyB-PIF light-inducible dimerization system together with the synPCB phycocyanobilin synthesis module. In the cited study, doxycycline treatment induced PCB synthesis and enabled PhyB-PIF light-inducible dimerization function.
Usefulness & Problems
Why this is useful
These vectors provide a delivery harness for stable introduction and inducible control of both an optogenetic dimerization module and its required chromophore biosynthesis pathway. This is useful when PhyB-PIF function must be coupled to regulated intracellular PCB production rather than constitutive expression.
Source:
These inducible lentiviral and transposon vectors were used to carry synPCB and PhyB-PIF. The abstract states that they enabled doxycycline-inducible PCB synthesis and PhyB-PIF LID activity.
Source:
stable delivery of synPCB and PhyB-PIF
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doxycycline-inducible activation of PCB synthesis and the LID system
Problem solved
The system addresses the need to co-deliver and drug-inducibly control PhyB-PIF and synPCB modules so that PCB synthesis and phytochrome-based light responses can be turned on by doxycycline. The supplied evidence does not show that the vectors themselves improve PCB synthesis beyond enabling inducible module expression.
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They provide a way to stably introduce and inducibly control both the chromophore synthesis module and the optogenetic dimerization system.
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enables inducible deployment of the chromophore synthesis system and optogenetic switch
Published Workflows
Objective: Improve genetically encoded phycocyanobilin synthesis to support phytochrome-based optogenetics and establish an inducible stable cell line containing the PhyB-PIF system.
Why it works: The workflow combines improved intracellular PCB production with inducible delivery of both the chromophore synthesis module and the PhyB-PIF switch, addressing the chromophore requirement that otherwise limits PhyB use outside organisms that naturally contain these tetrapyrroles.
genetically encoded PCB biosynthesisred/far-red light-inducible PhyB-PIF association and dissociationcomponent replacement with thermophilic cyanobacterial counterpartsfusion-protein engineeringpolycistronic expression designlentiviral and transposon vector incorporation
Stages
- 1.PCB synthesis module redesign(library_design)
This stage exists to improve intracellular PCB availability, which is required for PhyB-based optogenetic function.
Selection: Improve the previously developed synPCB system by changing component origins, protein architecture, and expression design.
- 2.Functional characterization of improved synPCB(functional_characterization)
This stage tests whether the redesign improved the intended output of the PCB synthesis system.
Selection: Measure PCB synthesis relative to the previous synPCB version.
- 3.Inducible vector-based stable cell line implementation(confirmatory_validation)
This stage confirms that the improved chromophore synthesis module and the optogenetic switch can be stably and inducibly deployed together in cells.
Selection: Incorporate PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors to enable doxycycline-inducible system deployment.
Steps
- 1.Replace PCB synthesis genes with thermophilic cyanobacterial counterpartsengineered PCB synthesis system
Redesign the synPCB module using alternative source genes for PCB synthesis.
The abstract presents this as the first engineering change in the improvement campaign.
- 2.Generate the tFnr-Fd chimeric proteinengineered component within synPCB
Modify electron transfer-related components by truncating Fnr and fusing it to Fd.
The abstract explicitly presents this as the second engineering modification after gene replacement.
- 3.Concatenate PCB synthesis genes with P2A peptide cDNAs for polycistronic expressionengineered PCB synthesis construct
Enable polycistronic expression of the redesigned PCB synthesis components.
The abstract presents polycistronic concatenation as the third design change after component replacement and chimera generation.
- 4.Assess PCB synthesis improvement relative to the previous versionengineered PCB synthesis system under test
Determine whether the redesigned synPCB increased PCB synthesis.
This measurement follows the design changes to test whether the intended improvement was achieved before stable deployment.
- 5.Incorporate synPCB and PhyB-PIF into drug inducible lentiviral and transposon vectorspayload systems and delivery harness
Prepare inducible delivery formats for stable cellular implementation of chromophore synthesis and the optogenetic switch.
This follows confirmation of improved PCB synthesis so the optimized module can be deployed together with PhyB-PIF in cells.
- 6.Induce PCB synthesis and the PhyB-PIF system with doxycyclineinducible deployed system
Confirm doxycycline-responsive induction of PCB synthesis and the PhyB-PIF LID system in the stable implementation.
Induction is tested after vector incorporation because doxycycline responsiveness depends on the inducible delivery setup.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A delivery strategy grouped with the mechanism branch because it determines how a system is instantiated and deployed in context.
Techniques
No technique tags yet.
Target processes
No target processes tagged yet.
Input: Light
Implementation Constraints
Implementation involved incorporating both PhyB-PIF and synPCB into drug-inducible lentiviral and transposon vectors. Operation required doxycycline treatment to induce PCB synthesis and the PhyB-PIF light-inducible dimerization system.
Evidence is limited to a single cited study and a brief statement that inducible delivery was achieved with lentiviral and transposon formats. The provided evidence does not report quantitative induction levels, cell-type scope, vector architecture, or comparative performance between the two delivery systems.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Observations
successMammalian Cell Lineapplication demo
Inferred from claim c3 during normalization. The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system. Derived from claim c3. Quoted text: successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
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successMammalian Cell Lineapplication demo
Inferred from claim c3 during normalization. The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system. Derived from claim c3. Quoted text: successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
Source:
successMammalian Cell Lineapplication demo
Inferred from claim c3 during normalization. The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system. Derived from claim c3. Quoted text: successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
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successMammalian Cell Lineapplication demo
Inferred from claim c3 during normalization. The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system. Derived from claim c3. Quoted text: successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
Source:
successMammalian Cell Lineapplication demo
Inferred from claim c3 during normalization. The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system. Derived from claim c3. Quoted text: successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
Source:
successMammalian Cell Lineapplication demo
Inferred from claim c3 during normalization. The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system. Derived from claim c3. Quoted text: successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
Source:
successMammalian Cell Lineapplication demo
Inferred from claim c3 during normalization. The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system. Derived from claim c3. Quoted text: successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
Source:
successMammalian Cell Lineapplication demo
Inferred from claim c3 during normalization. The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system. Derived from claim c3. Quoted text: successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
Source:
successMammalian Cell Lineapplication demo
Inferred from claim c3 during normalization. The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system. Derived from claim c3. Quoted text: successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
Source:
Supporting Sources
Ranked Claims
The PhyB-PIF light-inducible dimerization system controls both association and dissociation by light on the second time scale.
has a unique property of controlling both association and dissociation by light on the second time scale
response timescale second time scale
Drug inducible lentiviral and transposon vectors carrying PhyB-PIF and synPCB enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Drug inducible lentiviral and transposon vectors carrying PhyB-PIF and synPCB enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Drug inducible lentiviral and transposon vectors carrying PhyB-PIF and synPCB enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Drug inducible lentiviral and transposon vectors carrying PhyB-PIF and synPCB enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Drug inducible lentiviral and transposon vectors carrying PhyB-PIF and synPCB enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Drug inducible lentiviral and transposon vectors carrying PhyB-PIF and synPCB enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Drug inducible lentiviral and transposon vectors carrying PhyB-PIF and synPCB enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Drug inducible lentiviral and transposon vectors carrying PhyB-PIF and synPCB enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Drug inducible lentiviral and transposon vectors carrying PhyB-PIF and synPCB enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Incorporation of PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Incorporation of PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Incorporation of PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Incorporation of PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Incorporation of PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Incorporation of PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Incorporation of PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Concatenating the PCB synthesis genes with P2A peptide cDNAs for polycistronic expression resulted in an approximately 4-fold increase in PCB synthesis compared with the previous version.
these genes were concatenated with P2A peptide cDNAs for polycistronic expression, resulting in an approximately 4-fold increase in PCB synthesis compared with the previous version
PCB synthesis increase 4 fold
The PhyB-PIF red/far-red responsive light-inducible dimerization system requires a linear tetrapyrrole chromophore such as phytochromobilin or phycocyanobilin.
PhyB requires a linear tetrapyrrole chromophore such as phytochromobilin or phycocyanobilin (PCB)
The PhyB-PIF red/far-red responsive light-inducible dimerization system requires a linear tetrapyrrole chromophore such as phytochromobilin or phycocyanobilin.
PhyB requires a linear tetrapyrrole chromophore such as phytochromobilin or phycocyanobilin (PCB)
The PhyB-PIF red/far-red responsive light-inducible dimerization system requires a linear tetrapyrrole chromophore such as phytochromobilin or phycocyanobilin.
PhyB requires a linear tetrapyrrole chromophore such as phytochromobilin or phycocyanobilin (PCB)
The PhyB-PIF red/far-red responsive light-inducible dimerization system requires a linear tetrapyrrole chromophore such as phytochromobilin or phycocyanobilin.
PhyB requires a linear tetrapyrrole chromophore such as phytochromobilin or phycocyanobilin (PCB)
The PhyB-PIF red/far-red responsive light-inducible dimerization system requires a linear tetrapyrrole chromophore such as phytochromobilin or phycocyanobilin.
PhyB requires a linear tetrapyrrole chromophore such as phytochromobilin or phycocyanobilin (PCB)
The PhyB-PIF red/far-red responsive light-inducible dimerization system requires a linear tetrapyrrole chromophore such as phytochromobilin or phycocyanobilin.
PhyB requires a linear tetrapyrrole chromophore such as phytochromobilin or phycocyanobilin (PCB)
The PhyB-PIF red/far-red responsive light-inducible dimerization system requires a linear tetrapyrrole chromophore such as phytochromobilin or phycocyanobilin.
PhyB requires a linear tetrapyrrole chromophore such as phytochromobilin or phycocyanobilin (PCB)
The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The authors successfully established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The authors successfully established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The authors successfully established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The authors successfully established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The authors successfully established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The authors successfully established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The authors successfully established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
The improved synPCB design increased PCB synthesis by approximately 4-fold compared with the previous version.
resulting in an approximately 4-fold increase in PCB synthesis compared with the previous version
PCB synthesis increase versus previous version approximately 4-fold
The improved synPCB design increased PCB synthesis by approximately 4-fold compared with the previous version.
resulting in an approximately 4-fold increase in PCB synthesis compared with the previous version
PCB synthesis increase versus previous version approximately 4-fold
The improved synPCB design increased PCB synthesis by approximately 4-fold compared with the previous version.
resulting in an approximately 4-fold increase in PCB synthesis compared with the previous version
PCB synthesis increase versus previous version approximately 4-fold
Approval Evidence
1 source2 linked approval claimsfirst-pass slug drug-inducible-lentiviral-and-transposon-vectors
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Source:
inducible controlsupports
Drug inducible lentiviral and transposon vectors carrying PhyB-PIF and synPCB enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system expression or function.
we incorporated PhyB-PIF and synPCB into drug inducible lentiviral and transposon vectors, which enabled us to induce PCB synthesis and PhyB-PIF LID system by doxycycline treatment
Source:
system establishmentsupports
The authors established a stable cell line containing a genetically encoded PhyB-PIF light-inducible dimerization system.
successfully established a stable cell line containing a genetically encoded PhyB-PIF LID system
Source:
Comparisons
Source-backed strengths
The cited work states that these inducible lentiviral and transposon vectors enabled doxycycline-inducible PCB synthesis and PhyB-PIF light-inducible dimerization system activity. The associated PhyB-PIF system is described as controlling both association and dissociation by light on the second time scale.
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supports doxycycline-inducible control
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enabled establishment of a stable cell line
Ranked Citations