Source 1primary paper2022The FASEB Journal
Toolkit/intracellular spatially segregated reporters of optoPAK1 activity
intracellular spatially segregated reporters of optoPAK1 activity
Also known as: corresponding biosensor
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Intracellular spatially segregated reporters of optoPAK1 activity are genetically designed reporter constructs intended to report optoPAK1-dependent phosphorylation at defined intracellular locations. The available evidence indicates that they were created alongside the light-responsive PAK1 analog optoPAK1 to monitor its activity after light-triggered relocalization.
Usefulness & Problems
Why this is useful
These reporters are useful for assessing where within the cell optoPAK1 activity occurs after illumination-driven recruitment to specified intracellular sites. They address the need to couple optogenetic kinase targeting with location-resolved readouts of kinase action.
Problem solved
The tool helps solve the problem of measuring optoPAK1 activity in a spatially resolved manner inside cells rather than only inferring activity from bulk or nonlocalized outputs. The evidence supports intended use as intracellular reporters, but does not provide detailed quantitative benchmarking in the supplied material.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Techniques
Computational DesignTarget processes
No target processes tagged yet.
Input: Light
Implementation Constraints
Implementation appears to require co-expression with optoPAK1, a genetically expressed light-responsive PAK1 analog engineered for low dark-state activity and constitutive activity independent of endogenous regulation. The reporters are described only as intracellular spatially segregated constructs, so practical details such as targeting motifs, fluorophores, illumination parameters, and host system are not specified in the supplied evidence.
The supplied evidence does not describe reporter architecture, dynamic range, phosphorylation site sequence, temporal resolution, or validation across multiple compartments or cell types. Independent replication is not provided in the supplied material, and performance appears supported only by the originating study.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
OptoPAK1 was designed to function independently of endogenous biochemical regulation in a constitutively active manner with minimal activity in the dark state.
OptoPAK1 was designed to function independently of endogenous biochemical regulation in a constitutively active manner with minimal activity in the dark state.
OptoPAK1 was designed to function independently of endogenous biochemical regulation in a constitutively active manner with minimal activity in the dark state.
OptoPAK1 was designed to function independently of endogenous biochemical regulation in a constitutively active manner with minimal activity in the dark state.
OptoPAK1 was designed to function independently of endogenous biochemical regulation in a constitutively active manner with minimal activity in the dark state.
OptoPAK1 was designed to function independently of endogenous biochemical regulation in a constitutively active manner with minimal activity in the dark state.
OptoPAK1 was designed to function independently of endogenous biochemical regulation in a constitutively active manner with minimal activity in the dark state.
OptoPAK1 was designed to function independently of endogenous biochemical regulation in a constitutively active manner with minimal activity in the dark state.
OptoPAK1 was designed to function independently of endogenous biochemical regulation in a constitutively active manner with minimal activity in the dark state.
OptoPAK1 was designed to function independently of endogenous biochemical regulation in a constitutively active manner with minimal activity in the dark state.
OptoPAK1 was designed to function independently of endogenous biochemical regulation in a constitutively active manner with minimal activity in the dark state.
OptoPAK1 was designed to function independently of endogenous biochemical regulation in a constitutively active manner with minimal activity in the dark state.
OptoPAK1 was designed to function independently of endogenous biochemical regulation in a constitutively active manner with minimal activity in the dark state.
OptoPAK1 was designed to function independently of endogenous biochemical regulation in a constitutively active manner with minimal activity in the dark state.
The authors developed a genetically expressed, light-responsive optogenetic analog of PAK1 called optoPAK1.
We developed an engineering strategy to construct a genetically expressed, light-responsive optogenetic analog of PAK1 (optoPAK1)
The authors developed a genetically expressed, light-responsive optogenetic analog of PAK1 called optoPAK1.
We developed an engineering strategy to construct a genetically expressed, light-responsive optogenetic analog of PAK1 (optoPAK1)
The authors developed a genetically expressed, light-responsive optogenetic analog of PAK1 called optoPAK1.
We developed an engineering strategy to construct a genetically expressed, light-responsive optogenetic analog of PAK1 (optoPAK1)
The authors developed a genetically expressed, light-responsive optogenetic analog of PAK1 called optoPAK1.
We developed an engineering strategy to construct a genetically expressed, light-responsive optogenetic analog of PAK1 (optoPAK1)
The authors developed a genetically expressed, light-responsive optogenetic analog of PAK1 called optoPAK1.
We developed an engineering strategy to construct a genetically expressed, light-responsive optogenetic analog of PAK1 (optoPAK1)
The authors developed a genetically expressed, light-responsive optogenetic analog of PAK1 called optoPAK1.
We developed an engineering strategy to construct a genetically expressed, light-responsive optogenetic analog of PAK1 (optoPAK1)
The authors developed a genetically expressed, light-responsive optogenetic analog of PAK1 called optoPAK1.
We developed an engineering strategy to construct a genetically expressed, light-responsive optogenetic analog of PAK1 (optoPAK1)
Upon illumination, optoPAK1 migrates to specified intracellular sites.
upon illumination, optoPAK1 migrates to specified intracellular sites
Upon illumination, optoPAK1 migrates to specified intracellular sites.
upon illumination, optoPAK1 migrates to specified intracellular sites
Upon illumination, optoPAK1 migrates to specified intracellular sites.
upon illumination, optoPAK1 migrates to specified intracellular sites
Upon illumination, optoPAK1 migrates to specified intracellular sites.
upon illumination, optoPAK1 migrates to specified intracellular sites
Upon illumination, optoPAK1 migrates to specified intracellular sites.
upon illumination, optoPAK1 migrates to specified intracellular sites
Upon illumination, optoPAK1 migrates to specified intracellular sites.
upon illumination, optoPAK1 migrates to specified intracellular sites
Upon illumination, optoPAK1 migrates to specified intracellular sites.
upon illumination, optoPAK1 migrates to specified intracellular sites
The improved light-induced dimer system iLid was used to recruit and photoactivate the optoPAK1 protein analog at discrete subcellular domains.
We employed the improved light-induced dimer (iLid) system as a means to recruit and photoactivate the protein analog at discrete subcellular domains.
The improved light-induced dimer system iLid was used to recruit and photoactivate the optoPAK1 protein analog at discrete subcellular domains.
We employed the improved light-induced dimer (iLid) system as a means to recruit and photoactivate the protein analog at discrete subcellular domains.
The improved light-induced dimer system iLid was used to recruit and photoactivate the optoPAK1 protein analog at discrete subcellular domains.
We employed the improved light-induced dimer (iLid) system as a means to recruit and photoactivate the protein analog at discrete subcellular domains.
The improved light-induced dimer system iLid was used to recruit and photoactivate the optoPAK1 protein analog at discrete subcellular domains.
We employed the improved light-induced dimer (iLid) system as a means to recruit and photoactivate the protein analog at discrete subcellular domains.
The improved light-induced dimer system iLid was used to recruit and photoactivate the optoPAK1 protein analog at discrete subcellular domains.
We employed the improved light-induced dimer (iLid) system as a means to recruit and photoactivate the protein analog at discrete subcellular domains.
The improved light-induced dimer system iLid was used to recruit and photoactivate the optoPAK1 protein analog at discrete subcellular domains.
We employed the improved light-induced dimer (iLid) system as a means to recruit and photoactivate the protein analog at discrete subcellular domains.
The improved light-induced dimer system iLid was used to recruit and photoactivate the optoPAK1 protein analog at discrete subcellular domains.
We employed the improved light-induced dimer (iLid) system as a means to recruit and photoactivate the protein analog at discrete subcellular domains.
Preliminary data indicated that optoPAK1 phosphorylates the designed intracellular reporters in a light-dependent fashion.
preliminary data displayed that optoPAK1 phosphorylates these reporters in a light-dependent fashion
Preliminary data indicated that optoPAK1 phosphorylates the designed intracellular reporters in a light-dependent fashion.
preliminary data displayed that optoPAK1 phosphorylates these reporters in a light-dependent fashion
Preliminary data indicated that optoPAK1 phosphorylates the designed intracellular reporters in a light-dependent fashion.
preliminary data displayed that optoPAK1 phosphorylates these reporters in a light-dependent fashion
Preliminary data indicated that optoPAK1 phosphorylates the designed intracellular reporters in a light-dependent fashion.
preliminary data displayed that optoPAK1 phosphorylates these reporters in a light-dependent fashion
Preliminary data indicated that optoPAK1 phosphorylates the designed intracellular reporters in a light-dependent fashion.
preliminary data displayed that optoPAK1 phosphorylates these reporters in a light-dependent fashion
Preliminary data indicated that optoPAK1 phosphorylates the designed intracellular reporters in a light-dependent fashion.
preliminary data displayed that optoPAK1 phosphorylates these reporters in a light-dependent fashion
Preliminary data indicated that optoPAK1 phosphorylates the designed intracellular reporters in a light-dependent fashion.
preliminary data displayed that optoPAK1 phosphorylates these reporters in a light-dependent fashion
Approval Evidence
1 source1 linked approval claimfirst-pass slug intracellular-spatially-segregated-reporters-of-optopak1-activity
we designed intracellular spatially segregated reporters of optoPAK1 activity
Source:
reporter responsesupports
Preliminary data indicated that optoPAK1 phosphorylates the designed intracellular reporters in a light-dependent fashion.
preliminary data displayed that optoPAK1 phosphorylates these reporters in a light-dependent fashion
Source:
Comparisons
Source-backed strengths
A key strength is the explicit design of spatially segregated intracellular reporters matched to a genetically expressed, light-responsive PAK1 analog. Because optoPAK1 migrates to specified intracellular sites upon illumination, these reporters are positioned to detect activity in a subcellularly controlled context.
Source:
We developed an engineering strategy to construct a genetically expressed, light-responsive optogenetic analog of PAK1 (optoPAK1)
Ranked Citations
- 1.