Source 1primary paper2024Molecular Biology of the Cell
Toolkit/CRY2-CIB1 receptor optogenetic activation system
CRY2-CIB1 receptor optogenetic activation system
Also known as: CRY2-CIB1 bicistronic vector, optogenetic tool
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The CRY2-CIB1 receptor optogenetic activation system is a light-responsive multi-component switch based on the light-induced interaction between Cryptochrome 2 (CRY2) and CIB1. It was implemented as a bicistronic vector co-expressing CRY2 fused to the intracellular domain of a guidance receptor together with membrane-anchored CIB1, enabling local optical activation of receptor signaling in neurons.
Usefulness & Problems
Why this is useful
This system is useful for spatially restricted activation of receptor signaling in neurons, allowing local perturbation of growth cone behavior and actin-dependent remodeling. In the reported application, it was used to reorient growth cones and stimulate local actin polymerization for branch initiation along axonal shafts.
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this tool was used to reorient growth cones toward or away from the site of light activation and to stimulate local actin polymerization for branch initiation along axonal shafts
Source:
Here, we demonstrate the use of an optogenetic tool to activate receptor signaling locally to induce actin-mediated growth cone remodeling in neurons.
Problem solved
It addresses the problem of activating receptor intracellular signaling with subcellular spatial precision rather than global stimulation. The reported implementation enabled local activation of guidance receptor pathways to test how localized signaling controls growth cone dynamics in primary neurons.
Source:
this tool was used to reorient growth cones toward or away from the site of light activation and to stimulate local actin polymerization for branch initiation along axonal shafts
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
light-induced heterodimerizationlocal activation of receptor signalingmembrane recruitment of receptor intracellular domainsTechniques
No technique tags yet.
Target processes
recombinationsignalingInput: Light
Implementation Constraints
The reported construct design used a bicistronic vector to co-express CRY2 fused to the intracellular domain of a guidance receptor and a membrane-anchored CIB1. The evidence supports use in primary neurons, but the supplied material does not specify promoter choice, membrane anchor identity, illumination wavelength, or delivery method.
The supplied evidence is limited to a neuronal application study and does not establish performance across other cell types, receptors, or organisms. Quantitative information on activation kinetics, reversibility, light dose requirements, background activity, and independent replication is not provided in the supplied evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Observations
successPrimary Cellsapplication demoprimary neurons
Inferred from claim c1 during normalization. A CRY2-CIB1-based optogenetic tool enabled local activation of receptor signaling to induce actin-mediated growth cone remodeling in neurons. Derived from claim c1. Quoted text: Here, we demonstrate the use of an optogenetic tool to activate receptor signaling locally to induce actin-mediated growth cone remodeling in neurons.
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successPrimary Cellsapplication demoprimary neurons
Inferred from claim c2 during normalization. Activation of the PlexA4 receptor construct induced growth cone collapse in primary neurons. Derived from claim c2. Quoted text: When expressed in primary neurons, activation of the growth inhibitory PlexA4 receptor induced growth cone collapse
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successPrimary Cellsapplication demoprimary neurons
Inferred from claim c3 during normalization. Activation of the TrkA receptor construct increased growth cone size in primary neurons. Derived from claim c3. Quoted text: while activation of the growth stimulating TrkA receptor increased growth cone size
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successPrimary Cellsapplication demoprimary neurons
Inferred from claim c4 during normalization. Local activation of either receptor elicited the predicted response in light-activated growth cones and an opposite response in neighboring non-illuminated growth cones of the same neuron. Derived from claim c4. Quoted text: local activation of either receptor not only elicited the predicted response in light-activated growth cones but also an opposite response in neighboring no-light-exposed growth cones of the same neuron
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successPrimary Cellsapplication demoprimary neurons
Inferred from claim c5 during normalization. The optogenetic tool was used to reorient growth cones toward or away from the site of light activation and to stimulate local actin polymerization for branch initiation along axonal shafts. Derived from claim c5. Quoted text: this tool was used to reorient growth cones toward or away from the site of light activation and to stimulate local actin polymerization for branch initiation along axonal shafts
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Supporting Sources
Ranked Claims
The optogenetic tool was used to reorient growth cones toward or away from the site of light activation and to stimulate local actin polymerization for branch initiation along axonal shafts.
this tool was used to reorient growth cones toward or away from the site of light activation and to stimulate local actin polymerization for branch initiation along axonal shafts
Local activation of either receptor elicited the predicted response in light-activated growth cones and an opposite response in neighboring non-illuminated growth cones of the same neuron.
local activation of either receptor not only elicited the predicted response in light-activated growth cones but also an opposite response in neighboring no-light-exposed growth cones of the same neuron
Activation of the PlexA4 receptor construct induced growth cone collapse in primary neurons.
When expressed in primary neurons, activation of the growth inhibitory PlexA4 receptor induced growth cone collapse
Activation of the TrkA receptor construct increased growth cone size in primary neurons.
while activation of the growth stimulating TrkA receptor increased growth cone size
A CRY2-CIB1-based optogenetic tool enabled local activation of receptor signaling to induce actin-mediated growth cone remodeling in neurons.
Here, we demonstrate the use of an optogenetic tool to activate receptor signaling locally to induce actin-mediated growth cone remodeling in neurons.
Approval Evidence
1 source5 linked approval claimsfirst-pass slug cry2-cib1-receptor-optogenetic-activation-system
Based on the light-induced interaction between Cryptochrome 2 (CRY2) and CIB1, we generated a bicistronic vector to co-expresses CRY2 fused to the intracellular domain of a guidance receptor and a membrane-anchored CIB1.
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application demosupports
The optogenetic tool was used to reorient growth cones toward or away from the site of light activation and to stimulate local actin polymerization for branch initiation along axonal shafts.
this tool was used to reorient growth cones toward or away from the site of light activation and to stimulate local actin polymerization for branch initiation along axonal shafts
Source:
local nonlocal responsesupports
Local activation of either receptor elicited the predicted response in light-activated growth cones and an opposite response in neighboring non-illuminated growth cones of the same neuron.
local activation of either receptor not only elicited the predicted response in light-activated growth cones but also an opposite response in neighboring no-light-exposed growth cones of the same neuron
Source:
phenotypic effectsupports
Activation of the PlexA4 receptor construct induced growth cone collapse in primary neurons.
When expressed in primary neurons, activation of the growth inhibitory PlexA4 receptor induced growth cone collapse
Source:
phenotypic effectsupports
Activation of the TrkA receptor construct increased growth cone size in primary neurons.
while activation of the growth stimulating TrkA receptor increased growth cone size
Source:
tool capabilitysupports
A CRY2-CIB1-based optogenetic tool enabled local activation of receptor signaling to induce actin-mediated growth cone remodeling in neurons.
Here, we demonstrate the use of an optogenetic tool to activate receptor signaling locally to induce actin-mediated growth cone remodeling in neurons.
Source:
Comparisons
Source-backed strengths
The tool supported local receptor activation that produced distinct phenotypic outputs depending on the receptor intracellular domain used, including PlexA4-induced growth cone collapse and TrkA-associated growth cone enlargement in primary neurons. It also enabled opposite responses between illuminated and neighboring non-illuminated growth cones of the same neuron, indicating spatially resolved signaling effects.
Ranked Citations
- 1.
Derived from 5 linked claims and 5 validation observations. Example evidence: this tool was used to reorient growth cones toward or away from the site of light activation and to stimulate local actin polymerization for branch initiation along axonal shafts