Source 1primary paper2019
Toolkit/CRY2-CIBN optogenetic system
CRY2-CIBN optogenetic system
Also known as: CRY2-CIBN
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
CRY2-CIBN is a multi-component optogenetic switch used for acute, light-dependent recruitment of proteins to the plasma membrane. In the cited study, it was used to manipulate protein localization and interrogate Cdc42-dependent positive-feedback signaling.
Usefulness & Problems
Why this is useful
This system is useful for rapidly controlling subcellular protein localization with light, enabling perturbation of signaling events at the plasma membrane. In the cited work, it provided a way to test how localized recruitment of Cdc42 pathway components influences scaffold-mediated positive feedback and cell morphogenesis.
Problem solved
It addresses the problem of acutely and spatially manipulating protein localization to dissect causal relationships in polarity signaling. Specifically, it was used to probe how membrane recruitment of signaling components contributes to Cdc42 positive feedback and rod-shape formation.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
induced protein colocalizationlight-dependent protein recruitmentsubcellular relocalization to the plasma membraneTechniques
No technique tags yet.
Target processes
localizationsignalingInput: Light
Implementation Constraints
The study states that CRY2-CIBN was implemented for acute light-dependent protein recruitment to the plasma membrane. The provided evidence does not specify construct architecture, fusion orientation, illumination parameters, cofactors, or expression system details.
The supplied evidence documents plasma-membrane recruitment in one study context but does not provide quantitative performance metrics such as kinetics, reversibility, dynamic range, or wavelength dependence. Independent replication and validation across multiple organisms, targets, or delivery formats are not established by the provided evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Observations
successYeastapplication demoS. pombe
Inferred from claim c4 during normalization. Re-wired cells with restored positive feedback are viable and grow in a bipolar manner even when lacking otherwise essential Cdc42 activators. Derived from claim c4. Quoted text: Remarkably, such re-wired cells are viable and grow in a bipolar manner even when lacking otherwise essential Cdc42 activators.
Source:
Supporting Sources
Ranked Claims
An engineered interaction between the GEF and the Cdc42 effector Pak1 bypasses Scd2 function and restores positive feedback.
We show that Scd2 function is completely bypassed and positive feedback restored by an engineered interaction between the GEF and a Cdc42 effector, the Pak1 kinase.
The study implements the CRY2-CIBN optogenetic system for acute light-dependent protein recruitment to the plasma membrane.
Here, we implement the CRY2-CIBN optogenetic system for acute light-dependent protein recruitment to the plasma membrane
Optogenetic recruitment of constitutively active Cdc42 leads to co-recruitment of the GEF Scd1 in a manner dependent on the scaffold protein Scd2.
Indeed, optogenetic recruitment of constitutively active Cdc42 leads to co-recruitment of the GEF Scd1, in a manner dependent on the scaffold protein Scd2.
Ras1 GTPase plays a dual role in localizing and activating the GEF, thereby potentiating the feedback.
Interestingly, these cells reveal that Ras1 GTPase plays a dual role in localizing and activating the GEF, thus potentiating the feedback.
Scaffold-mediated positive feedback gated by Ras activity is minimally required for rod-shape formation.
We conclude that scaffold-mediated positive feedback, gated by Ras activity, is minimally required for rod-shape formation.
Re-wired cells with restored positive feedback are viable and grow in a bipolar manner even when lacking otherwise essential Cdc42 activators.
Remarkably, such re-wired cells are viable and grow in a bipolar manner even when lacking otherwise essential Cdc42 activators.
Approval Evidence
1 source6 linked approval claimsfirst-pass slug cry2-cibn-optogenetic-system
Here, we implement the CRY2-CIBN optogenetic system for acute light-dependent protein recruitment to the plasma membrane
Source:
engineering resultsupports
An engineered interaction between the GEF and the Cdc42 effector Pak1 bypasses Scd2 function and restores positive feedback.
We show that Scd2 function is completely bypassed and positive feedback restored by an engineered interaction between the GEF and a Cdc42 effector, the Pak1 kinase.
Source:
implementationsupports
The study implements the CRY2-CIBN optogenetic system for acute light-dependent protein recruitment to the plasma membrane.
Here, we implement the CRY2-CIBN optogenetic system for acute light-dependent protein recruitment to the plasma membrane
Source:
mechanistic effectsupports
Optogenetic recruitment of constitutively active Cdc42 leads to co-recruitment of the GEF Scd1 in a manner dependent on the scaffold protein Scd2.
Indeed, optogenetic recruitment of constitutively active Cdc42 leads to co-recruitment of the GEF Scd1, in a manner dependent on the scaffold protein Scd2.
Source:
mechanistic rolesupports
Ras1 GTPase plays a dual role in localizing and activating the GEF, thereby potentiating the feedback.
Interestingly, these cells reveal that Ras1 GTPase plays a dual role in localizing and activating the GEF, thus potentiating the feedback.
Source:
minimal requirementsupports
Scaffold-mediated positive feedback gated by Ras activity is minimally required for rod-shape formation.
We conclude that scaffold-mediated positive feedback, gated by Ras activity, is minimally required for rod-shape formation.
Source:
phenotypic resultsupports
Re-wired cells with restored positive feedback are viable and grow in a bipolar manner even when lacking otherwise essential Cdc42 activators.
Remarkably, such re-wired cells are viable and grow in a bipolar manner even when lacking otherwise essential Cdc42 activators.
Source:
Comparisons
Source-backed strengths
The reported implementation supports acute light-dependent recruitment to the plasma membrane, which is well suited for dynamic perturbation experiments. In the study, optogenetic recruitment of constitutively active Cdc42 led to Scd1 co-recruitment in a Scd2-dependent manner, and rewired cells with restored positive feedback were viable and grew in a bipolar manner.
Source:
We show that Scd2 function is completely bypassed and positive feedback restored by an engineered interaction between the GEF and a Cdc42 effector, the Pak1 kinase.
Ranked Citations
- 1.