Source 1primary paper2017Journal of Biotechnology
Toolkit/DMNP-EDTA-Cu system
DMNP-EDTA-Cu system
Also known as: DMNP-EDTA-Cu, photocaged Cu2+
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The DMNP-EDTA-Cu system is a light-activated, photocaged Cu2+ induction method for controlling transcription. Illumination releases Cu2+, which activates the Cu2+-responsive pCUP1 promoter from Saccharomyces cerevisiae to drive gene expression.
Usefulness & Problems
Why this is useful
This system enables optical control of a metal-inducible yeast promoter without direct constitutive addition of free Cu2+. It is useful for experiments requiring externally timed induction of transcription using light as the input signal.
Source:
This permits light-mediated control over the widely used Cu2+-inducible pCUP1 promoter from S. cerevisiae
Source:
we used the versatile photolabile chelator DMNP-EDTA ... to control Cu2+ release upon specific UV-A irradiation
Problem solved
It addresses the problem of achieving light-mediated activation of the S. cerevisiae pCUP1 promoter. Specifically, it converts illumination into release of Cu2+, the inducer required for pCUP1-dependent gene expression.
Source:
This permits light-mediated control over the widely used Cu2+-inducible pCUP1 promoter from S. cerevisiae
Published Workflows
Objective: Develop and screen a light-controlled yeast gene-expression system based on photocaged Cu2+ release to regulate a Cu2+-inducible promoter and optimize induction timing and expression output.
Why it works: The workflow couples a photolabile Cu2+ source to a Cu2+-responsive promoter, so light exposure controls inducer availability and thereby transcription. High-throughput per-well illumination and monitoring allow screening for induction timing and expression level.
UV-A-triggered photorelease of Cu2+ from DMNP-EDTACu2+-induced activation of the pCUP1 promotercomputational chemistry supporthigh-throughput online monitoringindividual well LED illumination
Stages
- 1.Computationally supported system design(library_design)
The abstract states that computational chemistry supported the choice or use of the photolabile chelator for Cu2+ release.
Selection: Use of DMNP-EDTA as a photolabile chelator to enable Cu2+ release upon UV-A irradiation.
- 2.High-throughput screening of induction conditions(broad_screen)
This stage was used to identify favorable induction timing and expression output for the DMNP-EDTA-Cu system.
Selection: Best induction time and expression level of eYFP under individual well illumination.
Steps
- 1.Use DMNP-EDTA as a photolabile chelator for Cu2+ photoreleaseengineered inducible system
Create a light-responsive Cu2+ source that can drive a Cu2+-inducible promoter.
A photoreleasable Cu2+ input is required before optical control of the pCUP1 promoter can be tested.
- 2.Screen induction time and eYFP expression with per-well LED illumination and online monitoringsystem under test and screening platform
Identify the best induction time and expression level for the photocaged Cu2+ expression system.
After establishing a light-responsive Cu2+ control system, screening is needed to optimize operational conditions and quantify expression behavior.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Target processes
recombinationselectionInput: Light
Implementation Constraints
The system is multi-component and requires the photocaged Cu2+ reagent DMNP-EDTA-Cu and a transcriptional output under control of the S. cerevisiae pCUP1 promoter. The available evidence indicates use in yeast, but does not provide construct architecture, illumination conditions, or delivery details in the supplied record.
The supplied evidence does not report quantitative induction levels, kinetics, reversibility, spectral parameters, or off-target effects. Validation is limited here to a single literature source describing light-controlled gene expression in yeast.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Photocaged Cu2+ release permits light-mediated control of the Cu2+-inducible pCUP1 promoter from S. cerevisiae.
This permits light-mediated control over the widely used Cu2+-inducible pCUP1 promoter from S. cerevisiae
Photocaged Cu2+ release permits light-mediated control of the Cu2+-inducible pCUP1 promoter from S. cerevisiae.
This permits light-mediated control over the widely used Cu2+-inducible pCUP1 promoter from S. cerevisiae
Photocaged Cu2+ release permits light-mediated control of the Cu2+-inducible pCUP1 promoter from S. cerevisiae.
This permits light-mediated control over the widely used Cu2+-inducible pCUP1 promoter from S. cerevisiae
Photocaged Cu2+ release permits light-mediated control of the Cu2+-inducible pCUP1 promoter from S. cerevisiae.
This permits light-mediated control over the widely used Cu2+-inducible pCUP1 promoter from S. cerevisiae
Photocaged Cu2+ release permits light-mediated control of the Cu2+-inducible pCUP1 promoter from S. cerevisiae.
This permits light-mediated control over the widely used Cu2+-inducible pCUP1 promoter from S. cerevisiae
Photocaged Cu2+ release permits light-mediated control of the Cu2+-inducible pCUP1 promoter from S. cerevisiae.
This permits light-mediated control over the widely used Cu2+-inducible pCUP1 promoter from S. cerevisiae
Photocaged Cu2+ release permits light-mediated control of the Cu2+-inducible pCUP1 promoter from S. cerevisiae.
This permits light-mediated control over the widely used Cu2+-inducible pCUP1 promoter from S. cerevisiae
Photocaged Cu2+ release permits light-mediated control of the Cu2+-inducible pCUP1 promoter from S. cerevisiae.
This permits light-mediated control over the widely used Cu2+-inducible pCUP1 promoter from S. cerevisiae
Caged Cu2+-based optical expression regulation provides minimally invasive, easy-to-control, parallel, automated, temporal, and quantitative control and is presented as a beneficial alternative to induction by pipetting CuCl2.
we realized a minimally invasive, easy-to-control, parallel and automated optical expression regulation via caged Cu2+ allowing temporal and quantitative control as a beneficial alternative to conventional induction via pipetting CuCl2 as effector molecule
Caged Cu2+-based optical expression regulation provides minimally invasive, easy-to-control, parallel, automated, temporal, and quantitative control and is presented as a beneficial alternative to induction by pipetting CuCl2.
we realized a minimally invasive, easy-to-control, parallel and automated optical expression regulation via caged Cu2+ allowing temporal and quantitative control as a beneficial alternative to conventional induction via pipetting CuCl2 as effector molecule
Caged Cu2+-based optical expression regulation provides minimally invasive, easy-to-control, parallel, automated, temporal, and quantitative control and is presented as a beneficial alternative to induction by pipetting CuCl2.
we realized a minimally invasive, easy-to-control, parallel and automated optical expression regulation via caged Cu2+ allowing temporal and quantitative control as a beneficial alternative to conventional induction via pipetting CuCl2 as effector molecule
Caged Cu2+-based optical expression regulation provides minimally invasive, easy-to-control, parallel, automated, temporal, and quantitative control and is presented as a beneficial alternative to induction by pipetting CuCl2.
we realized a minimally invasive, easy-to-control, parallel and automated optical expression regulation via caged Cu2+ allowing temporal and quantitative control as a beneficial alternative to conventional induction via pipetting CuCl2 as effector molecule
Caged Cu2+-based optical expression regulation provides minimally invasive, easy-to-control, parallel, automated, temporal, and quantitative control and is presented as a beneficial alternative to induction by pipetting CuCl2.
we realized a minimally invasive, easy-to-control, parallel and automated optical expression regulation via caged Cu2+ allowing temporal and quantitative control as a beneficial alternative to conventional induction via pipetting CuCl2 as effector molecule
Caged Cu2+-based optical expression regulation provides minimally invasive, easy-to-control, parallel, automated, temporal, and quantitative control and is presented as a beneficial alternative to induction by pipetting CuCl2.
we realized a minimally invasive, easy-to-control, parallel and automated optical expression regulation via caged Cu2+ allowing temporal and quantitative control as a beneficial alternative to conventional induction via pipetting CuCl2 as effector molecule
Caged Cu2+-based optical expression regulation provides minimally invasive, easy-to-control, parallel, automated, temporal, and quantitative control and is presented as a beneficial alternative to induction by pipetting CuCl2.
we realized a minimally invasive, easy-to-control, parallel and automated optical expression regulation via caged Cu2+ allowing temporal and quantitative control as a beneficial alternative to conventional induction via pipetting CuCl2 as effector molecule
Caged Cu2+-based optical expression regulation provides minimally invasive, easy-to-control, parallel, automated, temporal, and quantitative control and is presented as a beneficial alternative to induction by pipetting CuCl2.
we realized a minimally invasive, easy-to-control, parallel and automated optical expression regulation via caged Cu2+ allowing temporal and quantitative control as a beneficial alternative to conventional induction via pipetting CuCl2 as effector molecule
The study reports the first example of a caged metal ion used to regulate recombinant gene expression.
thus constitutes the first example of a caged metal ion to regulate recombinant gene expression
The study reports the first example of a caged metal ion used to regulate recombinant gene expression.
thus constitutes the first example of a caged metal ion to regulate recombinant gene expression
The study reports the first example of a caged metal ion used to regulate recombinant gene expression.
thus constitutes the first example of a caged metal ion to regulate recombinant gene expression
The study reports the first example of a caged metal ion used to regulate recombinant gene expression.
thus constitutes the first example of a caged metal ion to regulate recombinant gene expression
The study reports the first example of a caged metal ion used to regulate recombinant gene expression.
thus constitutes the first example of a caged metal ion to regulate recombinant gene expression
The study reports the first example of a caged metal ion used to regulate recombinant gene expression.
thus constitutes the first example of a caged metal ion to regulate recombinant gene expression
The study reports the first example of a caged metal ion used to regulate recombinant gene expression.
thus constitutes the first example of a caged metal ion to regulate recombinant gene expression
The study reports the first example of a caged metal ion used to regulate recombinant gene expression.
thus constitutes the first example of a caged metal ion to regulate recombinant gene expression
The DMNP-EDTA-Cu system was screened for optimal induction time and eYFP expression level using a high-throughput online monitoring system with individual well illumination.
We screened our novel DMNP-EDTA-Cu system for best induction time and expression level of eYFP with a high-throughput online monitoring system equipped with an LED array for individual illumination of every single well
The DMNP-EDTA-Cu system was screened for optimal induction time and eYFP expression level using a high-throughput online monitoring system with individual well illumination.
We screened our novel DMNP-EDTA-Cu system for best induction time and expression level of eYFP with a high-throughput online monitoring system equipped with an LED array for individual illumination of every single well
The DMNP-EDTA-Cu system was screened for optimal induction time and eYFP expression level using a high-throughput online monitoring system with individual well illumination.
We screened our novel DMNP-EDTA-Cu system for best induction time and expression level of eYFP with a high-throughput online monitoring system equipped with an LED array for individual illumination of every single well
The DMNP-EDTA-Cu system was screened for optimal induction time and eYFP expression level using a high-throughput online monitoring system with individual well illumination.
We screened our novel DMNP-EDTA-Cu system for best induction time and expression level of eYFP with a high-throughput online monitoring system equipped with an LED array for individual illumination of every single well
The DMNP-EDTA-Cu system was screened for optimal induction time and eYFP expression level using a high-throughput online monitoring system with individual well illumination.
We screened our novel DMNP-EDTA-Cu system for best induction time and expression level of eYFP with a high-throughput online monitoring system equipped with an LED array for individual illumination of every single well
The DMNP-EDTA-Cu system was screened for optimal induction time and eYFP expression level using a high-throughput online monitoring system with individual well illumination.
We screened our novel DMNP-EDTA-Cu system for best induction time and expression level of eYFP with a high-throughput online monitoring system equipped with an LED array for individual illumination of every single well
The DMNP-EDTA-Cu system was screened for optimal induction time and eYFP expression level using a high-throughput online monitoring system with individual well illumination.
We screened our novel DMNP-EDTA-Cu system for best induction time and expression level of eYFP with a high-throughput online monitoring system equipped with an LED array for individual illumination of every single well
The DMNP-EDTA-Cu system was screened for optimal induction time and eYFP expression level using a high-throughput online monitoring system with individual well illumination.
We screened our novel DMNP-EDTA-Cu system for best induction time and expression level of eYFP with a high-throughput online monitoring system equipped with an LED array for individual illumination of every single well
DMNP-EDTA can be used to control Cu2+ release upon specific UV-A irradiation.
we used the versatile photolabile chelator DMNP-EDTA ... to control Cu2+ release upon specific UV-A irradiation
DMNP-EDTA can be used to control Cu2+ release upon specific UV-A irradiation.
we used the versatile photolabile chelator DMNP-EDTA ... to control Cu2+ release upon specific UV-A irradiation
DMNP-EDTA can be used to control Cu2+ release upon specific UV-A irradiation.
we used the versatile photolabile chelator DMNP-EDTA ... to control Cu2+ release upon specific UV-A irradiation
DMNP-EDTA can be used to control Cu2+ release upon specific UV-A irradiation.
we used the versatile photolabile chelator DMNP-EDTA ... to control Cu2+ release upon specific UV-A irradiation
DMNP-EDTA can be used to control Cu2+ release upon specific UV-A irradiation.
we used the versatile photolabile chelator DMNP-EDTA ... to control Cu2+ release upon specific UV-A irradiation
DMNP-EDTA can be used to control Cu2+ release upon specific UV-A irradiation.
we used the versatile photolabile chelator DMNP-EDTA ... to control Cu2+ release upon specific UV-A irradiation
DMNP-EDTA can be used to control Cu2+ release upon specific UV-A irradiation.
we used the versatile photolabile chelator DMNP-EDTA ... to control Cu2+ release upon specific UV-A irradiation
DMNP-EDTA can be used to control Cu2+ release upon specific UV-A irradiation.
we used the versatile photolabile chelator DMNP-EDTA ... to control Cu2+ release upon specific UV-A irradiation
Approval Evidence
1 source5 linked approval claimsfirst-pass slug dmnp-edta-cu-system
We screened our novel DMNP-EDTA-Cu system for best induction time and expression level of eYFP ... allowing temporal and quantitative control as a beneficial alternative to conventional induction via pipetting CuCl2 as effector molecule.
Source:
applicationsupports
Photocaged Cu2+ release permits light-mediated control of the Cu2+-inducible pCUP1 promoter from S. cerevisiae.
This permits light-mediated control over the widely used Cu2+-inducible pCUP1 promoter from S. cerevisiae
Source:
comparative advantagesupports
Caged Cu2+-based optical expression regulation provides minimally invasive, easy-to-control, parallel, automated, temporal, and quantitative control and is presented as a beneficial alternative to induction by pipetting CuCl2.
we realized a minimally invasive, easy-to-control, parallel and automated optical expression regulation via caged Cu2+ allowing temporal and quantitative control as a beneficial alternative to conventional induction via pipetting CuCl2 as effector molecule
Source:
noveltysupports
The study reports the first example of a caged metal ion used to regulate recombinant gene expression.
thus constitutes the first example of a caged metal ion to regulate recombinant gene expression
Source:
screening resultsupports
The DMNP-EDTA-Cu system was screened for optimal induction time and eYFP expression level using a high-throughput online monitoring system with individual well illumination.
We screened our novel DMNP-EDTA-Cu system for best induction time and expression level of eYFP with a high-throughput online monitoring system equipped with an LED array for individual illumination of every single well
Source:
tool capabilitysupports
DMNP-EDTA can be used to control Cu2+ release upon specific UV-A irradiation.
we used the versatile photolabile chelator DMNP-EDTA ... to control Cu2+ release upon specific UV-A irradiation
Source:
Comparisons
Source-backed strengths
The reported strength is direct coupling of light exposure to activation of a defined Cu2+-responsive promoter. The available evidence supports its application for light-controlled gene expression in yeast, but provides limited broader performance detail here.
Source:
we realized a minimally invasive, easy-to-control, parallel and automated optical expression regulation via caged Cu2+ allowing temporal and quantitative control as a beneficial alternative to conventional induction via pipetting CuCl2 as effector molecule
Source:
thus constitutes the first example of a caged metal ion to regulate recombinant gene expression
Ranked Citations
- 1.