Source 1primary paper2019
Toolkit/TIP-LITer
TIP-LITer
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
TIP-LITer is a light-inducible TetR-based gene circuit component in which TetR is fused to a Tet-inhibitory peptide through the LOV2 photosensory domain. The reported design places inhibitory peptide control under light-sensitive regulation in mammalian cells.
Usefulness & Problems
Why this is useful
This tool is useful for introducing optical control into TetR-based regulatory circuits by coupling TetR to a Tet-inhibitory peptide via LOV2. The available evidence supports its use as part of the LITer optogenetic toolset in mammalian cells, but does not provide detailed performance data in the supplied source.
Problem solved
TIP-LITer addresses the problem of making TetR circuit activity responsive to light by embedding a Tet-inhibitory peptide within a LOV2-linked fusion architecture. The supplied evidence indicates that it was designed as a light-sensitive variant of a TetR regulatory module, but does not further specify the exact experimental bottleneck solved.
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
Degradationinhibitory peptide-mediated repression tuninglight-dependent allosteric switchingTechniques
No technique tags yet.
Target processes
degradationInput: Light
Implementation Constraints
The reported construct consists of TetR fused to a Tet-inhibitory peptide through the LOV2 light-sensitive domain. The source identifies the LITer toolset as operating in mammalian cells, but does not provide construct orientation details, linker design, illumination parameters, or delivery method in the supplied evidence.
The supplied evidence is limited to composition and does not report quantitative light responses, dynamic range, kinetics, wavelength dependence, or benchmarking against alternative systems. Independent replication is not provided in the supplied 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 tip-liter
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) ... 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 Tet-inhibiting peptide (TIP) ... 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 key strength is its modular composition: TetR, a Tet-inhibitory peptide, and the LOV2 light-sensitive domain are combined in a single fusion design. The source also places this construct within a mammalian-cell optogenetic circuit context, supporting its intended compatibility with mammalian expression systems.
Ranked Citations
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