Source 1primary paper2016Journal of Cell Science
Toolkit/Opto-PIP3
Opto-PIP3
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Opto-PIP3 is an optogenetic construct used to elevate or spatially pattern PI(3,4,5)P3 signaling in cells. In adipocytes, its activation largely mimicked the maximal effect of insulin on IRAP-pHluorin translocation, supporting the sufficiency of PI3K-generated PIP3 for much of this membrane-trafficking response.
Usefulness & Problems
Why this is useful
This tool is useful for dissecting the contribution of PI3K-generated PI(3,4,5)P3 to insulin-responsive membrane trafficking. The reported adipocyte experiments indicate that it can separate PIP3-driven effects from downstream Akt-dependent signaling and reveal Akt-independent components of GLUT4-related translocation.
Source:
Here, we describe new optogenetic tools based on CRY2 and the N-terminus of CIB1 (CIBN). We used these 'Opto' modules to activate PI3K and Akt selectively in time and space in 3T3-L1 adipocytes.
Problem solved
It addresses the problem of testing whether PI3K/PIP3 signaling is sufficient to drive insulin-responsive vesicle translocation, rather than merely correlating with it during receptor stimulation. The cited study used it to distinguish roles of PI3K/PIP3 and Akt in adipocyte insulin action.
Problem links
Need inducible protein relocalization or recruitment
DerivedOpto-PIP3 is an optogenetic construct used to elevate or spatially pattern PI(3,4,5)P3 signaling in cells. In adipocytes, its activation largely mimicked the maximal effect of insulin on IRAP-pHluorin translocation, supporting the sufficiency of PI3K-generated PIP3 for much of this membrane-trafficking response.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Mechanisms
optogenetic control of pi(3,4,5)p3 signalingsubcellular localization/patterning of signaling activityTechniques
No technique tags yet.
Target processes
localizationInput: Chemical
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: regulator
The available evidence indicates use in adipocytes with IRAP-pHluorin translocation as the functional readout. Practical details such as the optogenetic module, chromophore requirements, construct design, expression strategy, and stimulation wavelength are not provided in the supplied evidence.
The supplied evidence is limited to a single study and a specific adipocyte trafficking assay centered on IRAP-pHluorin translocation. The evidence provided does not specify molecular architecture, illumination parameters, kinetics, dynamic range, or validation in additional cell types or in vivo settings.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Opto-PIP3 largely mimicked the maximal effects of insulin stimulation on IRAP-pHluorin translocation, whereas Opto-Akt only partially triggered translocation.
Strikingly, Opto-PIP3 largely mimicked the maximal effects of insulin stimulation, whereas Opto-Akt only partially triggered translocation.
Opto-PIP3 largely mimicked the maximal effects of insulin stimulation on IRAP-pHluorin translocation, whereas Opto-Akt only partially triggered translocation.
Strikingly, Opto-PIP3 largely mimicked the maximal effects of insulin stimulation, whereas Opto-Akt only partially triggered translocation.
Opto-PIP3 largely mimicked the maximal effects of insulin stimulation on IRAP-pHluorin translocation, whereas Opto-Akt only partially triggered translocation.
Strikingly, Opto-PIP3 largely mimicked the maximal effects of insulin stimulation, whereas Opto-Akt only partially triggered translocation.
Opto-PIP3 largely mimicked the maximal effects of insulin stimulation on IRAP-pHluorin translocation, whereas Opto-Akt only partially triggered translocation.
Strikingly, Opto-PIP3 largely mimicked the maximal effects of insulin stimulation, whereas Opto-Akt only partially triggered translocation.
Opto-PIP3 largely mimicked the maximal effects of insulin stimulation on IRAP-pHluorin translocation, whereas Opto-Akt only partially triggered translocation.
Strikingly, Opto-PIP3 largely mimicked the maximal effects of insulin stimulation, whereas Opto-Akt only partially triggered translocation.
Opto-PIP3 largely mimicked the maximal effects of insulin stimulation on IRAP-pHluorin translocation, whereas Opto-Akt only partially triggered translocation.
Strikingly, Opto-PIP3 largely mimicked the maximal effects of insulin stimulation, whereas Opto-Akt only partially triggered translocation.
Opto-PIP3 largely mimicked the maximal effects of insulin stimulation on IRAP-pHluorin translocation, whereas Opto-Akt only partially triggered translocation.
Strikingly, Opto-PIP3 largely mimicked the maximal effects of insulin stimulation, whereas Opto-Akt only partially triggered translocation.
Drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3.
Conversely, drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3
Drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3.
Conversely, drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3
Drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3.
Conversely, drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3
Drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3.
Conversely, drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3
Drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3.
Conversely, drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3
Drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3.
Conversely, drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3
Drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3.
Conversely, drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3
PI3K and Akt play distinct roles in adipocyte insulin action, and PI3K stimulates Akt-independent pathways important for GLUT4 translocation.
Taken together, these results indicate that PI3K and Akt play distinct roles, and that PI3K stimulates Akt-independent pathways that are important for GLUT4 translocation.
PI3K and Akt play distinct roles in adipocyte insulin action, and PI3K stimulates Akt-independent pathways important for GLUT4 translocation.
Taken together, these results indicate that PI3K and Akt play distinct roles, and that PI3K stimulates Akt-independent pathways that are important for GLUT4 translocation.
PI3K and Akt play distinct roles in adipocyte insulin action, and PI3K stimulates Akt-independent pathways important for GLUT4 translocation.
Taken together, these results indicate that PI3K and Akt play distinct roles, and that PI3K stimulates Akt-independent pathways that are important for GLUT4 translocation.
PI3K and Akt play distinct roles in adipocyte insulin action, and PI3K stimulates Akt-independent pathways important for GLUT4 translocation.
Taken together, these results indicate that PI3K and Akt play distinct roles, and that PI3K stimulates Akt-independent pathways that are important for GLUT4 translocation.
PI3K and Akt play distinct roles in adipocyte insulin action, and PI3K stimulates Akt-independent pathways important for GLUT4 translocation.
Taken together, these results indicate that PI3K and Akt play distinct roles, and that PI3K stimulates Akt-independent pathways that are important for GLUT4 translocation.
PI3K and Akt play distinct roles in adipocyte insulin action, and PI3K stimulates Akt-independent pathways important for GLUT4 translocation.
Taken together, these results indicate that PI3K and Akt play distinct roles, and that PI3K stimulates Akt-independent pathways that are important for GLUT4 translocation.
PI3K and Akt play distinct roles in adipocyte insulin action, and PI3K stimulates Akt-independent pathways important for GLUT4 translocation.
Taken together, these results indicate that PI3K and Akt play distinct roles, and that PI3K stimulates Akt-independent pathways that are important for GLUT4 translocation.
Focal targeting of Akt to a region of the cell marked sites where IRAP-pHluorin vesicles fused, supporting local Akt-mediated regulation of exocytosis.
In spatial optogenetic studies, focal targeting of Akt to a region of the cell marked the sites where IRAP-pHluorin vesicles fused, supporting the idea that local Akt-mediated signaling regulates exocytosis.
Focal targeting of Akt to a region of the cell marked sites where IRAP-pHluorin vesicles fused, supporting local Akt-mediated regulation of exocytosis.
In spatial optogenetic studies, focal targeting of Akt to a region of the cell marked the sites where IRAP-pHluorin vesicles fused, supporting the idea that local Akt-mediated signaling regulates exocytosis.
Focal targeting of Akt to a region of the cell marked sites where IRAP-pHluorin vesicles fused, supporting local Akt-mediated regulation of exocytosis.
In spatial optogenetic studies, focal targeting of Akt to a region of the cell marked the sites where IRAP-pHluorin vesicles fused, supporting the idea that local Akt-mediated signaling regulates exocytosis.
Focal targeting of Akt to a region of the cell marked sites where IRAP-pHluorin vesicles fused, supporting local Akt-mediated regulation of exocytosis.
In spatial optogenetic studies, focal targeting of Akt to a region of the cell marked the sites where IRAP-pHluorin vesicles fused, supporting the idea that local Akt-mediated signaling regulates exocytosis.
Focal targeting of Akt to a region of the cell marked sites where IRAP-pHluorin vesicles fused, supporting local Akt-mediated regulation of exocytosis.
In spatial optogenetic studies, focal targeting of Akt to a region of the cell marked the sites where IRAP-pHluorin vesicles fused, supporting the idea that local Akt-mediated signaling regulates exocytosis.
Focal targeting of Akt to a region of the cell marked sites where IRAP-pHluorin vesicles fused, supporting local Akt-mediated regulation of exocytosis.
In spatial optogenetic studies, focal targeting of Akt to a region of the cell marked the sites where IRAP-pHluorin vesicles fused, supporting the idea that local Akt-mediated signaling regulates exocytosis.
Focal targeting of Akt to a region of the cell marked sites where IRAP-pHluorin vesicles fused, supporting local Akt-mediated regulation of exocytosis.
In spatial optogenetic studies, focal targeting of Akt to a region of the cell marked the sites where IRAP-pHluorin vesicles fused, supporting the idea that local Akt-mediated signaling regulates exocytosis.
The study describes optogenetic tools based on CRY2 and CIBN that selectively activate PI3K and Akt in time and space in 3T3-L1 adipocytes.
Here, we describe new optogenetic tools based on CRY2 and the N-terminus of CIB1 (CIBN). We used these 'Opto' modules to activate PI3K and Akt selectively in time and space in 3T3-L1 adipocytes.
The study describes optogenetic tools based on CRY2 and CIBN that selectively activate PI3K and Akt in time and space in 3T3-L1 adipocytes.
Here, we describe new optogenetic tools based on CRY2 and the N-terminus of CIB1 (CIBN). We used these 'Opto' modules to activate PI3K and Akt selectively in time and space in 3T3-L1 adipocytes.
The study describes optogenetic tools based on CRY2 and CIBN that selectively activate PI3K and Akt in time and space in 3T3-L1 adipocytes.
Here, we describe new optogenetic tools based on CRY2 and the N-terminus of CIB1 (CIBN). We used these 'Opto' modules to activate PI3K and Akt selectively in time and space in 3T3-L1 adipocytes.
The study describes optogenetic tools based on CRY2 and CIBN that selectively activate PI3K and Akt in time and space in 3T3-L1 adipocytes.
Here, we describe new optogenetic tools based on CRY2 and the N-terminus of CIB1 (CIBN). We used these 'Opto' modules to activate PI3K and Akt selectively in time and space in 3T3-L1 adipocytes.
The study describes optogenetic tools based on CRY2 and CIBN that selectively activate PI3K and Akt in time and space in 3T3-L1 adipocytes.
Here, we describe new optogenetic tools based on CRY2 and the N-terminus of CIB1 (CIBN). We used these 'Opto' modules to activate PI3K and Akt selectively in time and space in 3T3-L1 adipocytes.
The study describes optogenetic tools based on CRY2 and CIBN that selectively activate PI3K and Akt in time and space in 3T3-L1 adipocytes.
Here, we describe new optogenetic tools based on CRY2 and the N-terminus of CIB1 (CIBN). We used these 'Opto' modules to activate PI3K and Akt selectively in time and space in 3T3-L1 adipocytes.
The study describes optogenetic tools based on CRY2 and CIBN that selectively activate PI3K and Akt in time and space in 3T3-L1 adipocytes.
Here, we describe new optogenetic tools based on CRY2 and the N-terminus of CIB1 (CIBN). We used these 'Opto' modules to activate PI3K and Akt selectively in time and space in 3T3-L1 adipocytes.
Approval Evidence
1 source3 linked approval claimsfirst-pass slug opto-pip3
Strikingly, Opto-PIP3 largely mimicked the maximal effects of insulin stimulation
Source:
comparative effectsupports
Opto-PIP3 largely mimicked the maximal effects of insulin stimulation on IRAP-pHluorin translocation, whereas Opto-Akt only partially triggered translocation.
Strikingly, Opto-PIP3 largely mimicked the maximal effects of insulin stimulation, whereas Opto-Akt only partially triggered translocation.
Source:
inhibitor interactionsupports
Drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3.
Conversely, drug-mediated inhibition of Akt only partially dampened the translocation response of Opto-PIP3
Source:
mechanistic conclusionsupports
PI3K and Akt play distinct roles in adipocyte insulin action, and PI3K stimulates Akt-independent pathways important for GLUT4 translocation.
Taken together, these results indicate that PI3K and Akt play distinct roles, and that PI3K stimulates Akt-independent pathways that are important for GLUT4 translocation.
Source:
Comparisons
Source-backed strengths
In the cited adipocyte context, Opto-PIP3 largely reproduced the maximal insulin effect on IRAP-pHluorin translocation, whereas Opto-Akt only partially triggered translocation. Akt inhibition only partially dampened the Opto-PIP3 response, further supporting the conclusion that PIP3 engages Akt-independent pathways important for GLUT4 translocation.
Source:
Strikingly, Opto-PIP3 largely mimicked the maximal effects of insulin stimulation, whereas Opto-Akt only partially triggered translocation.
Compared with PROTAC
Opto-PIP3 and PROTAC address a similar problem space because they share localization.
Shared frame: same top-level item type; shared target processes: localization; same primary input modality: chemical
Compared with Ra1-based Ras-E2F1 anchoring aptamer
Opto-PIP3 and Ra1-based Ras-E2F1 anchoring aptamer address a similar problem space because they share localization.
Shared frame: same top-level item type; shared target processes: localization; same primary input modality: chemical
Compared with Ra1-based Ras-p65 anchoring aptamer
Opto-PIP3 and Ra1-based Ras-p65 anchoring aptamer address a similar problem space because they share localization.
Shared frame: same top-level item type; shared target processes: localization; same primary input modality: chemical
Ranked Citations
- 1.