Source 1primary paper2015ACS Synthetic Biology
Toolkit/synthetic transcription factor
synthetic transcription factor
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The synthetic transcription factor is an engineered cargo used with a red/far-red light-regulated phytochrome-based nuclear localization system. In mammalian cells and zebrafish, its light-controlled nuclear translocation was used to drive transgene expression as a functional readout of regulated protein import.
Usefulness & Problems
Why this is useful
This construct is useful as a downstream effector that converts optically controlled nuclear import into transcriptional output. The reported application shows that patterned light can regulate nuclear import and associated target protein activity in mammalian cells and zebrafish.
Source:
This engineered transcription factor is used as a cargo whose nuclear entry can be controlled to drive transgene expression. The abstract presents it as a downstream functional demonstration of the light-regulated localization system.
Source:
light-controlled transgene expression
Problem solved
It addresses the problem of linking light-regulated nuclear localization to a measurable gene-expression outcome. Specifically, it enables transgene expression to be controlled by induced nuclear entry of a synthetic transcription factor.
Source:
It provides a way to convert light-controlled nuclear import into a gene expression output.
Source:
couples light-controlled nuclear translocation to transcriptional output
Problem links
couples light-controlled nuclear translocation to transcriptional output
LiteratureIt provides a way to convert light-controlled nuclear import into a gene expression output.
Source:
It provides a way to convert light-controlled nuclear import into a gene expression output.
Published Workflows
Red Light-Regulated Reversible Nuclear Localization of Proteins in Mammalian Cells and Zebrafish
2015Objective: Develop and apply a reversible red/far-red light-controlled system to regulate nuclear localization and downstream activity of proteins in vertebrate contexts.
Why it works: The workflow is based on reconstructing a red light-dependent phytochrome B/PIF3 nuclear import mechanism in a nonplant environment and then using that principle to control nuclear import and activity of target proteins.
red light-dependent phytochrome B and phytochrome-interacting factor 3 mediated nuclear importlight-controlled translocation of target proteins into the nucleussynthetic reconstruction in a nonplant environmentspatiotemporal projection of light patterns
Stages
- 1.Mechanism reconstruction and validation in a nonplant environment(functional_characterization)
This stage establishes the underlying import principle before applying it to regulate target proteins.
Selection: Reconstruction and validation of red light-dependent Arabidopsis phytochrome B nuclear import mediated by phytochrome-interacting factor 3 outside plants.
- 2.Light-patterned regulation of target protein nuclear import and activity(secondary_characterization)
This stage tests whether the reconstructed mechanism can be used as a practical control modality for target proteins.
Selection: Use of spatiotemporal light patterns to regulate nuclear import and activity of target proteins.
- 3.Functional demonstration in mammalian cells and zebrafish(confirmatory_validation)
This stage confirms that light-controlled nuclear translocation can produce a downstream gene-expression effect in vertebrate systems.
Selection: Translocation of a synthetic transcription factor into the nucleus to drive transgene expression in mammalian cells and zebrafish.
Steps
- 1.Synthetically reconstruct phytochrome B/PIF3-mediated nuclear import in a nonplant environment
Port the Arabidopsis red light-dependent nuclear import mechanism into a nonplant setting.
The abstract states this was done first to establish the principle underlying the later control system.
- 2.Validate the reconstructed import mechanism
Confirm that the reconstructed system supports red light-dependent nuclear import.
Validation follows reconstruction so the authors can use the established principle for downstream regulation experiments.
- 3.Apply spatiotemporal light patterns to regulate target protein nuclear import and activitylight-regulated control system
Use the validated principle to control target protein localization and activity with spatial and temporal precision.
The abstract explicitly says this was done on the basis of the previously established principle.
- 4.Translocate a synthetic transcription factor into the nucleus to drive transgene expressionregulated cargo and control system
Demonstrate that light-controlled nuclear localization can produce a downstream transcriptional output in vertebrate systems.
This follows mechanistic and control demonstrations to confirm functional utility in mammalian cells and zebrafish.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Mechanisms
light-controlled nuclear localizationreversible photoswitching by red and far-red lighttranscriptional activation following nuclear translocationTechniques
No technique tags yet.
Target processes
transcriptionImplementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: sensor
Use requires expression of the synthetic transcription factor together with the light-regulated nuclear localization system in mammalian cells or zebrafish. The evidence supports use as a fused or engineered cargo whose nuclear entry is optically controlled, but it does not provide construct sequence, domain composition, or delivery details.
The available evidence does not specify the transcription factor architecture, activation domain, DNA-binding module, promoter design, or quantitative performance. The evidence also does not show that the construct itself confers localization control independently of the accompanying phytochrome-based system.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
Using this principle, the authors regulated nuclear import and activity of target proteins by spatiotemporal projection of light patterns.
On the basis of this principle we next regulated nuclear import and activity of target proteins by the spatiotemporal projection of light patterns.
The data demonstrate the first in vivo application of a plant phytochrome-based optogenetic tool in vertebrates.
These data demonstrate the first in vivo application of a plant phytochrome-based optogenetic tool in vertebrates
The paper reports a red light-inducible and far-red light-reversible synthetic system for controlling nuclear localization of proteins in mammalian cells and zebrafish.
Here we report the development of a red light-inducible and far-red light-reversible synthetic system for controlling nuclear localization of proteins in mammalian cells and zebrafish.
Approval Evidence
1 source1 linked approval claimfirst-pass slug synthetic-transcription-factor
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
Source:
application demosupports
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression.
Source:
Comparisons
Source-stated alternatives
No specific alternative transcriptional control constructs are named in the abstract.
Source:
No specific alternative transcriptional control constructs are named in the abstract.
Source-backed strengths
The construct was demonstrated as functional cargo in both mammalian cells and zebrafish, indicating use across cultured cells and an animal model. Its activity is coupled to spatiotemporal light control of nuclear import, enabling regulation of transgene expression through controlled nuclear translocation.
Source:
used in mammalian cells and zebrafish
Compared with DNA scaffolding
synthetic transcription factor and DNA scaffolding address a similar problem space because they share transcription.
Shared frame: same top-level item type; shared target processes: transcription
Strengths here: looks easier to implement in practice.
Compared with genetically engineered biosensors
synthetic transcription factor and genetically engineered biosensors address a similar problem space because they share transcription.
Shared frame: same top-level item type; shared target processes: transcription
Compared with synthetic promoters
synthetic transcription factor and synthetic promoters address a similar problem space because they share transcription.
Shared frame: same top-level item type; shared target processes: transcription
Relative tradeoffs: appears more independently replicated; looks easier to implement in practice.
Ranked Citations
- 1.