Source 1primary paper2017Journal of Biotechnology
Toolkit/pCUP1 promoter expression module
pCUP1 promoter expression module
Also known as: Cu2+-inducible pCUP1 promoter, pCUP1
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The pCUP1 promoter expression module is a light-addressable gene expression system in Saccharomyces cerevisiae that couples photocaged Cu2+ to the native Cu2+-inducible pCUP1 promoter. Illumination releases Cu2+, which activates pCUP1-driven recombinant gene expression.
Usefulness & Problems
Why this is useful
This module enables optical control of a native yeast metal-responsive promoter without direct manual addition of soluble CuCl2 at the time of induction. The cited study presents this approach as minimally invasive and compatible with parallel, automated, temporal, and quantitative regulation of gene expression.
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 controlling Cu2+-inducible pCUP1 expression with light rather than by pipetting CuCl2 into cultures. This provides a route to externally timed induction of recombinant gene expression in yeast using optical input.
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.
Mechanisms
light-triggered metal ion releasemetal-ion-dependent transcriptional inductionPhotocleavageTechniques
No technique tags yet.
Target processes
No target processes tagged yet.
Input: Light
Implementation Constraints
Implementation requires the native Cu2+-responsive pCUP1 promoter from Saccharomyces cerevisiae and a photocaged Cu2+ source that releases Cu2+ upon illumination. The evidence supports promoter activation by light-triggered Cu2+ uncaging, but does not provide construct architecture, illumination parameters, or strain-specific deployment details.
The supplied evidence is limited to a single source describing the concept and its claimed advantages. No quantitative performance metrics, wavelength requirements, dynamic range, leakiness, toxicity limits, or independent replication are provided in the evidence set.
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 source1 linked approval claimfirst-pass slug pcup1-promoter-expression-module
This permits light-mediated control over the widely used Cu2+-inducible pCUP1 promoter from S. cerevisiae.
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:
Comparisons
Source-backed strengths
The reported system permits light-mediated activation of the Cu2+-inducible pCUP1 promoter in S. cerevisiae through photocaged Cu2+ release. The source specifically highlights ease of control and potential advantages for minimally invasive, parallel, automated, temporal, and quantitative expression regulation.
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.