Source 1primary paper2013ACS Synthetic Biology
Toolkit/cryptochrome 2-mediated optogenetic C-RAF activation tool
cryptochrome 2-mediated optogenetic C-RAF activation tool
Also known as: light-activated C-RAF, optogenetic tool
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The cryptochrome 2-mediated optogenetic C-RAF activation tool is a mammalian-cell optogenetic system in which Arabidopsis thaliana cryptochrome 2 drives blue light-dependent dimerization to activate the protein kinase C-RAF. It enables reversible optical control of C-RAF signaling outputs, including ERK1/2 phosphorylation and serum response factor-mediated gene expression.
Usefulness & Problems
Why this is useful
This tool provides optical control over protein kinase activity, allowing modulation of signaling strength and duration with blue light. It is useful for interrogating C-RAF-dependent signaling dynamics while separating C-RAF activation from upstream surface receptor inputs.
Source:
This optogenetic tool enables reversible control of protein kinase activity in signal duration and strength.
Source:
Mediated by Arabidopsis thaliana photoreceptor cryptochrome 2, we activated the protein kinase C-RAF by blue light-dependent dimerization
Problem solved
It addresses the problem of activating C-RAF in mammalian cells without relying on endogenous receptor-triggered upstream signaling events. The system also enables controlled study of how C-RAF activation kinetics and dose influence downstream ERK1/2 phosphorylation and transcriptional output.
Problem links
Need conditional control of signaling activity
DerivedThe cryptochrome 2-mediated optogenetic C-RAF activation tool is a mammalian-cell optogenetic system in which Arabidopsis thaliana cryptochrome 2 enables blue light-dependent dimerization to activate the protein kinase C-RAF. It provides reversible optical control of C-RAF signaling output, including ERK1/2 phosphorylation and serum response factor-mediated gene expression.
Need precise spatiotemporal control with light input
DerivedThe cryptochrome 2-mediated optogenetic C-RAF activation tool is a mammalian-cell optogenetic system in which Arabidopsis thaliana cryptochrome 2 enables blue light-dependent dimerization to activate the protein kinase C-RAF. It provides reversible optical control of C-RAF signaling output, including ERK1/2 phosphorylation and serum response factor-mediated gene expression.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Techniques
No technique tags yet.
Target processes
signalingInput: Light
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: multi component delivery burdenimplementation constraint: spectral hardware requirementoperating role: regulatorswitch architecture: multi componentswitch architecture: recruitment
The system is mediated by Arabidopsis thaliana cryptochrome 2 and operates through blue light-dependent dimerization in mammalian cells. The available evidence supports a domain-fusion optogenetic design, but specific fusion partners, illumination parameters, and delivery or expression details are not provided in the supplied material.
The supplied evidence is limited to a single source study in mammalian cells and does not provide quantitative performance metrics, construct architecture, or validation across multiple cell types or organisms. No information is provided here on phototoxicity, basal activity, dynamic range, or long-term expression effects.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Light-activated C-RAF controls serum response factor-mediated gene expression.
Additionally, light-activated C-RAF controls serum response factor-mediated gene expression.
Light-induced heterodimerization of C-RAF with a kinase-dead B-RAF mutant demonstrates that B-RAF enhances C-RAF activity as a scaffold.
Light-induced heterodimerization of C-RAF with a kinase-dead mutant of B-RAF demonstrates the enhancing role of B-RAF as a scaffold for C-RAF activity
Light activation of C-RAF is fast, reversible, and time- and dose-dependent as monitored by ERK1/2 phosphorylation.
The activation by light is fast, reversible, and not only time but also dose dependent as monitored by phosphorylation of ERK1/2.
Blue light-dependent dimerization of C-RAF allows decoupling of C-RAF activation from upstream signaling events induced by surface receptors.
allowing for decoupling from upstream signaling events induced by surface receptors
This optogenetic tool enables reversible control of protein kinase activity in signal duration and strength.
This optogenetic tool enables reversible control of protein kinase activity in signal duration and strength.
A cryptochrome 2-mediated optogenetic tool activates C-RAF by blue light-dependent dimerization in mammalian cells.
Mediated by Arabidopsis thaliana photoreceptor cryptochrome 2, we activated the protein kinase C-RAF by blue light-dependent dimerization
Approval Evidence
1 source6 linked approval claimsfirst-pass slug cryptochrome-2-mediated-optogenetic-c-raf-activation-tool
Mediated by Arabidopsis thaliana photoreceptor cryptochrome 2, we activated the protein kinase C-RAF by blue light-dependent dimerization
Source:
downstream effectsupports
Light-activated C-RAF controls serum response factor-mediated gene expression.
Additionally, light-activated C-RAF controls serum response factor-mediated gene expression.
Source:
mechanistic inferencesupports
Light-induced heterodimerization of C-RAF with a kinase-dead B-RAF mutant demonstrates that B-RAF enhances C-RAF activity as a scaffold.
Light-induced heterodimerization of C-RAF with a kinase-dead mutant of B-RAF demonstrates the enhancing role of B-RAF as a scaffold for C-RAF activity
Source:
response propertysupports
Light activation of C-RAF is fast, reversible, and time- and dose-dependent as monitored by ERK1/2 phosphorylation.
The activation by light is fast, reversible, and not only time but also dose dependent as monitored by phosphorylation of ERK1/2.
Source:
signaling decouplingsupports
Blue light-dependent dimerization of C-RAF allows decoupling of C-RAF activation from upstream signaling events induced by surface receptors.
allowing for decoupling from upstream signaling events induced by surface receptors
Source:
tool capabilitysupports
This optogenetic tool enables reversible control of protein kinase activity in signal duration and strength.
This optogenetic tool enables reversible control of protein kinase activity in signal duration and strength.
Source:
tool functionsupports
A cryptochrome 2-mediated optogenetic tool activates C-RAF by blue light-dependent dimerization in mammalian cells.
Mediated by Arabidopsis thaliana photoreceptor cryptochrome 2, we activated the protein kinase C-RAF by blue light-dependent dimerization
Source:
Comparisons
Source-backed strengths
Reported performance includes fast, reversible, time-dependent, and dose-dependent activation as monitored by ERK1/2 phosphorylation. The tool also controls serum response factor-mediated gene expression and was used to show that light-induced heterodimerization with a kinase-dead B-RAF mutant can enhance C-RAF activity through a scaffold effect.
Compared with fusion proteins with large N-terminal anchors
cryptochrome 2-mediated optogenetic C-RAF activation tool and fusion proteins with large N-terminal anchors address a similar problem space because they share signaling.
Shared frame: same top-level item type; shared target processes: signaling; shared mechanisms: heterodimerization; same primary input modality: light
Compared with LOVpep/ePDZb
cryptochrome 2-mediated optogenetic C-RAF activation tool and LOVpep/ePDZb address a similar problem space because they share signaling.
Shared frame: same top-level item type; shared target processes: signaling; shared mechanisms: heterodimerization; same primary input modality: light
Relative tradeoffs: appears more independently replicated; looks easier to implement in practice.
Compared with tandem-dimer nano (tdnano)
cryptochrome 2-mediated optogenetic C-RAF activation tool and tandem-dimer nano (tdnano) address a similar problem space because they share signaling.
Shared frame: same top-level item type; shared target processes: signaling; shared mechanisms: heterodimerization; same primary input modality: light
Ranked Citations
- 1.