Source 1primary paper2019
Toolkit/Deg-LITer
Deg-LITer
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Deg-LITer is a multi-component optogenetic gene circuit in which the TetR repressor is fused to a degradation tag through the LOV2 light-sensitive domain. It is part of the LITer toolset for light-controlled regulation in mammalian cells.
Usefulness & Problems
Why this is useful
This design is useful as a light-responsive means to couple a transcriptional regulator to a degradation module in mammalian cells. The available evidence supports its role as a component of an optogenetic regulation platform, but does not provide performance details in the supplied source.
Problem solved
Deg-LITer addresses the need for light-controlled regulation of gene circuits in mammalian cells using a TetR-based architecture linked to a degradation tag. The supplied evidence does not further specify the exact experimental bottleneck or comparative advantage it was designed to overcome.
Published Workflows
Objective: Engineer optogenetic negative-feedback gene circuits in mammalian cells that reduce gene expression noise while enabling precise control of expression and downstream phenotypic perturbation.
Why it works: The workflow combines optogenetic control with negative-feedback repression, which the abstract frames as a route to lower expression noise while retaining tunable control in mammalian cells.
negative feedbacklight-responsive control through LOV2TetR-based repressiongene circuit engineeringoptogenetic controlcomparative performance evaluationapplication to oncogene perturbation
Stages
- 1.LITer circuit design and build(library_design)
This stage creates the LITer toolset architectures that are later evaluated for performance and application.
Selection: Construct optogenetic negative-feedback circuits using TetR fused with TIP or a degradation tag through LOV2.
- 2.Performance characterization against existing optogenetic systems(functional_characterization)
This stage establishes whether the engineered LITer circuits outperform prior optogenetic systems on the targeted control and noise axes.
Selection: Measure gene expression control range and noise reduction relative to existing optogenetic systems.
- 3.Application to KRAS(G12V) perturbation(confirmatory_validation)
This stage tests whether the LITer architecture is useful beyond reporter control by applying it to an oncogenic payload and downstream phenotype readouts.
Selection: Use the LITer architecture to control KRAS(G12V) expression and assess downstream phospho-ERK and proliferation effects.
Objective: Engineer optogenetic negative-feedback gene circuits in mammalian cells to achieve reduced gene expression noise and precise control of expression, then apply the architecture to perturb KRAS(G12V) signaling outputs.
Why it works: The abstract states that the circuits use TetR fused to either a Tet-inhibiting peptide or a degradation tag through LOV2, coupling light sensitivity to negative-feedback repression in order to tune expression and reduce noise.
genetic negative feedbacklight-sensitive control through LOV2TetR-mediated repressionoptogenetic circuit engineering
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
Degradationlight-regulated degradationlov2-based photosensory controlnegative-feedback gene regulationTechniques
No technique tags yet.
Target processes
degradationInput: Light
Implementation Constraints
The reported construct contains TetR, a degradation tag, and the LOV2 light-sensitive domain in a fusion architecture. It is described in the context of mammalian cells, but the source excerpt does not specify construct orientation, promoter design, illumination conditions, or required cofactors.
The supplied evidence is limited to circuit composition and does not report activation wavelength, dynamic range, kinetics, leakiness, or target gene outputs. Independent replication and breadth of validation cannot be established from the provided material.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Source 2primary paper2019Nucleic Acids Research
Ranked Claims
The LITer gene circuit architecture was used to control expression of KRAS(G12V) and study downstream phospho-ERK levels and cellular proliferation.
Moreover, we use the LITer gene circuit architecture to control gene expression of the cancer oncogene KRAS(G12V) and study its downstream effects through phospho-ERK levels and cellular proliferation.
The LITer gene circuit architecture was used to control expression of KRAS(G12V) and study downstream phospho-ERK levels and cellular proliferation.
Moreover, we use the LITer gene circuit architecture to control gene expression of the cancer oncogene KRAS(G12V) and study its downstream effects through phospho-ERK levels and cellular proliferation.
The LITer gene circuit architecture was used to control expression of KRAS(G12V) and study downstream phospho-ERK levels and cellular proliferation.
Moreover, we use the LITer gene circuit architecture to control gene expression of the cancer oncogene KRAS(G12V) and study its downstream effects through phospho-ERK levels and cellular proliferation.
The LITer gene circuit architecture was used to control expression of KRAS(G12V) and study downstream phospho-ERK levels and cellular proliferation.
Moreover, we use the LITer gene circuit architecture to control gene expression of the cancer oncogene KRAS(G12V) and study its downstream effects through phospho-ERK levels and cellular proliferation.
The LITer gene circuit architecture was used to control expression of KRAS(G12V) and study downstream phospho-ERK levels and cellular proliferation.
Moreover, we use the LITer gene circuit architecture to control gene expression of the cancer oncogene KRAS(G12V) and study its downstream effects through phospho-ERK levels and cellular proliferation.
The LITer gene circuit architecture was used to control KRAS(G12V) expression and examine downstream phospho-ERK levels and cellular proliferation.
Moreover, we use the LITer gene circuit architecture to control gene expression of the cancer oncogene KRAS(G12V) and study its downstream effects through phospho-ERK levels and cellular proliferation.
The LITer gene circuit architecture was used to control KRAS(G12V) expression and examine downstream phospho-ERK levels and cellular proliferation.
Moreover, we use the LITer gene circuit architecture to control gene expression of the cancer oncogene KRAS(G12V) and study its downstream effects through phospho-ERK levels and cellular proliferation.
The LITer gene circuit architecture was used to control KRAS(G12V) expression and examine downstream phospho-ERK levels and cellular proliferation.
Moreover, we use the LITer gene circuit architecture to control gene expression of the cancer oncogene KRAS(G12V) and study its downstream effects through phospho-ERK levels and cellular proliferation.
The LITer gene circuit architecture was used to control KRAS(G12V) expression and examine downstream phospho-ERK levels and cellular proliferation.
Moreover, we use the LITer gene circuit architecture to control gene expression of the cancer oncogene KRAS(G12V) and study its downstream effects through phospho-ERK levels and cellular proliferation.
The LITer gene circuit architecture was used to control KRAS(G12V) expression and examine downstream phospho-ERK levels and cellular proliferation.
Moreover, we use the LITer gene circuit architecture to control gene expression of the cancer oncogene KRAS(G12V) and study its downstream effects through phospho-ERK levels and cellular proliferation.
The LITer toolset uses TetR fused with either a Tet-Inhibitory peptide or a degradation tag through the LOV2 light-sensitive domain.
We build a toolset of these noise-reducing Light-Inducible Tuner (LITer) gene circuits using the TetR repressor fused with a Tet-Inhibitory peptide (TIP) or a degradation tag through the light-sensitive LOV2 protein domain.
The LITer toolset uses TetR fused with either a Tet-Inhibitory peptide or a degradation tag through the LOV2 light-sensitive domain.
We build a toolset of these noise-reducing Light-Inducible Tuner (LITer) gene circuits using the TetR repressor fused with a Tet-Inhibitory peptide (TIP) or a degradation tag through the light-sensitive LOV2 protein domain.
The LITer toolset uses TetR fused with either a Tet-Inhibitory peptide or a degradation tag through the LOV2 light-sensitive domain.
We build a toolset of these noise-reducing Light-Inducible Tuner (LITer) gene circuits using the TetR repressor fused with a Tet-Inhibitory peptide (TIP) or a degradation tag through the light-sensitive LOV2 protein domain.
The LITer toolset uses TetR fused with either a Tet-Inhibitory peptide or a degradation tag through the LOV2 light-sensitive domain.
We build a toolset of these noise-reducing Light-Inducible Tuner (LITer) gene circuits using the TetR repressor fused with a Tet-Inhibitory peptide (TIP) or a degradation tag through the light-sensitive LOV2 protein domain.
The LITer toolset uses TetR fused with either a Tet-Inhibitory peptide or a degradation tag through the LOV2 light-sensitive domain.
We build a toolset of these noise-reducing Light-Inducible Tuner (LITer) gene circuits using the TetR repressor fused with a Tet-Inhibitory peptide (TIP) or a degradation tag through the light-sensitive LOV2 protein domain.
The authors engineered optogenetic negative-feedback gene circuits in mammalian cells for noise-reduced precise gene expression control.
Here, we engineer optogenetic negative-feedback gene circuits in mammalian cells to achieve noise-reduction for precise gene expression control.
The authors engineered optogenetic negative-feedback gene circuits in mammalian cells for noise-reduced precise gene expression control.
Here, we engineer optogenetic negative-feedback gene circuits in mammalian cells to achieve noise-reduction for precise gene expression control.
The authors engineered optogenetic negative-feedback gene circuits in mammalian cells for noise-reduced precise gene expression control.
Here, we engineer optogenetic negative-feedback gene circuits in mammalian cells to achieve noise-reduction for precise gene expression control.
The authors engineered optogenetic negative-feedback gene circuits in mammalian cells for noise-reduced precise gene expression control.
Here, we engineer optogenetic negative-feedback gene circuits in mammalian cells to achieve noise-reduction for precise gene expression control.
The authors engineered optogenetic negative-feedback gene circuits in mammalian cells for noise-reduced precise gene expression control.
Here, we engineer optogenetic negative-feedback gene circuits in mammalian cells to achieve noise-reduction for precise gene expression control.
LITer circuits provide nearly 4-fold gene expression control.
These LITers provide a range of nearly 4-fold gene expression control
gene expression control range 4 fold
LITer circuits provide nearly 4-fold gene expression control.
These LITers provide a range of nearly 4-fold gene expression control
gene expression control range 4 fold
LITer circuits provide nearly 4-fold gene expression control.
These LITers provide a range of nearly 4-fold gene expression control
gene expression control range 4 fold
LITer circuits provide nearly 4-fold gene expression control.
These LITers provide a range of nearly 4-fold gene expression control
gene expression control range 4 fold
LITer circuits provide nearly 4-fold gene expression control.
These LITers provide a range of nearly 4-fold gene expression control
gene expression control range 4 fold
LITer circuits provide nearly 4-fold gene expression control and up to five-fold noise reduction relative to existing optogenetic systems.
These LITers provide nearly a range of 4-fold gene expression control and up to five-fold noise reduction from existing optogenetic systems.
gene expression control range 4 foldnoise reduction versus existing optogenetic systems 5 fold
LITer circuits provide nearly 4-fold gene expression control and up to five-fold noise reduction relative to existing optogenetic systems.
These LITers provide nearly a range of 4-fold gene expression control and up to five-fold noise reduction from existing optogenetic systems.
gene expression control range 4 foldnoise reduction versus existing optogenetic systems 5 fold
LITer circuits provide nearly 4-fold gene expression control and up to five-fold noise reduction relative to existing optogenetic systems.
These LITers provide nearly a range of 4-fold gene expression control and up to five-fold noise reduction from existing optogenetic systems.
gene expression control range 4 foldnoise reduction versus existing optogenetic systems 5 fold
LITer circuits provide nearly 4-fold gene expression control and up to five-fold noise reduction relative to existing optogenetic systems.
These LITers provide nearly a range of 4-fold gene expression control and up to five-fold noise reduction from existing optogenetic systems.
gene expression control range 4 foldnoise reduction versus existing optogenetic systems 5 fold
LITer circuits provide nearly 4-fold gene expression control and up to five-fold noise reduction relative to existing optogenetic systems.
These LITers provide nearly a range of 4-fold gene expression control and up to five-fold noise reduction from existing optogenetic systems.
gene expression control range 4 foldnoise reduction versus existing optogenetic systems 5 fold
LITer circuits reduce gene expression noise by up to 5-fold relative to existing optogenetic systems.
and up to 5-fold noise reduction from existing optogenetic systems
noise reduction 5 fold
LITer circuits reduce gene expression noise by up to 5-fold relative to existing optogenetic systems.
and up to 5-fold noise reduction from existing optogenetic systems
noise reduction 5 fold
LITer circuits reduce gene expression noise by up to 5-fold relative to existing optogenetic systems.
and up to 5-fold noise reduction from existing optogenetic systems
noise reduction 5 fold
LITer circuits reduce gene expression noise by up to 5-fold relative to existing optogenetic systems.
and up to 5-fold noise reduction from existing optogenetic systems
noise reduction 5 fold
LITer circuits reduce gene expression noise by up to 5-fold relative to existing optogenetic systems.
and up to 5-fold noise reduction from existing optogenetic systems
noise reduction 5 fold
LITer gene circuits enable optogenetic negative-feedback control of gene expression in mammalian cells.
Here, we engineer optogenetic gene circuits into mammalian cells to achieve noise-reduction for precise gene expression control by genetic, in vitro negative feedback.
LITer gene circuits enable optogenetic negative-feedback control of gene expression in mammalian cells.
Here, we engineer optogenetic gene circuits into mammalian cells to achieve noise-reduction for precise gene expression control by genetic, in vitro negative feedback.
LITer gene circuits enable optogenetic negative-feedback control of gene expression in mammalian cells.
Here, we engineer optogenetic gene circuits into mammalian cells to achieve noise-reduction for precise gene expression control by genetic, in vitro negative feedback.
LITer gene circuits enable optogenetic negative-feedback control of gene expression in mammalian cells.
Here, we engineer optogenetic gene circuits into mammalian cells to achieve noise-reduction for precise gene expression control by genetic, in vitro negative feedback.
LITer gene circuits enable optogenetic negative-feedback control of gene expression in mammalian cells.
Here, we engineer optogenetic gene circuits into mammalian cells to achieve noise-reduction for precise gene expression control by genetic, in vitro negative feedback.
LITer optogenetic platforms should enable precise spatiotemporal perturbations for studying multicellular phenotypes in developmental biology, oncology, and other biomedical research fields.
Overall, these novel LITer optogenetic platforms should enable precise spatiotemporal perturbations for studying multicellular phenotypes in developmental biology, oncology, and other biomedical fields of research.
LITer optogenetic platforms should enable precise spatiotemporal perturbations for studying multicellular phenotypes in developmental biology, oncology, and other biomedical research fields.
Overall, these novel LITer optogenetic platforms should enable precise spatiotemporal perturbations for studying multicellular phenotypes in developmental biology, oncology, and other biomedical fields of research.
LITer optogenetic platforms should enable precise spatiotemporal perturbations for studying multicellular phenotypes in developmental biology, oncology, and other biomedical research fields.
Overall, these novel LITer optogenetic platforms should enable precise spatiotemporal perturbations for studying multicellular phenotypes in developmental biology, oncology, and other biomedical fields of research.
LITer optogenetic platforms should enable precise spatiotemporal perturbations for studying multicellular phenotypes in developmental biology, oncology, and other biomedical research fields.
Overall, these novel LITer optogenetic platforms should enable precise spatiotemporal perturbations for studying multicellular phenotypes in developmental biology, oncology, and other biomedical fields of research.
LITer optogenetic platforms should enable precise spatiotemporal perturbations for studying multicellular phenotypes in developmental biology, oncology, and other biomedical research fields.
Overall, these novel LITer optogenetic platforms should enable precise spatiotemporal perturbations for studying multicellular phenotypes in developmental biology, oncology, and other biomedical fields of research.
Approval Evidence
2 sources1 linked approval claimfirst-pass slug deg-liter
We build a toolset of these noise-reducing Light-Inducible Tuner (LITer) gene circuits using the TetR repressor fused with ... a degradation tag through the light-sensitive LOV2 protein domain.
Source:
We build a toolset of these noise-reducing Light-Inducible Tuner (LITer) gene circuits using the TetR repressor fused with ... a degradation tag through the light-sensitive LOV2 protein domain.
Source:
compositionsupports
The LITer toolset uses TetR fused with either a Tet-Inhibitory peptide or a degradation tag through the LOV2 light-sensitive domain.
We build a toolset of these noise-reducing Light-Inducible Tuner (LITer) gene circuits using the TetR repressor fused with a Tet-Inhibitory peptide (TIP) or a degradation tag through the light-sensitive LOV2 protein domain.
Source:
Comparisons
Source-backed strengths
A clear strength is its modular composition: TetR is connected to a degradation tag via the LOV2 photosensory domain, enabling a genetically encoded light-responsive design. The source also places it within a broader LITer toolset for mammalian optogenetics, but no quantitative validation is provided here.
Ranked Citations
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