Source 1review2014Current Protocols in Cell Biology
Toolkit/Cry2/CIB
Cry2/CIB
Also known as: Cryptochrome 2 (Cry2)/CIB
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Cry2/CIB is a genetically encoded blue-light-activated protein dimerization module derived from Arabidopsis thaliana. It is used to optically induce protein-protein interactions and has been applied to control transcription, protein localization, protein secretion, and, when coupled to BAX, light-triggered apoptosis.
Usefulness & Problems
Why this is useful
This module provides noninvasive optical control over protein association, enabling temporal regulation of cellular processes with light. The cited literature presents CRY2/CIB as a practical optical dimerizer for manipulating transcription, localization, and secretion, and as a component of a light-activated apoptotic actuator.
Source:
Previously, we have employed Cryptochrome 2 (Cry2)/CIB, a blue light photoreceptor protein - protein dimerization module from A. thaliana in conjunction with BAX, an OMM targeting pro-apoptotic protein, for light-mediated initiation of mitochondrial outer membrane permeabilization (MOMP) and downstream apoptosis.
Source:
We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8, for use in controlling transcription, protein localization, and protein secretion using light.
Source:
we provide instructions and software for constructing a pulse-controlled LED device for use in experiments requiring extended light treatments
Problem solved
Cry2/CIB addresses the problem of controlling intracellular protein interactions on demand without constitutive association. In the apoptosis application, optimization specifically targeted reduction of light-independent cell death and improved experimental control by altering photophysical properties of the Cry2/CIB interaction.
Source:
We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8, for use in controlling transcription, protein localization, and protein secretion using light.
Problem links
Need conditional recombination or state switching
DerivedCry2/CIB is a genetically encoded blue-light-activated protein dimerization module derived from Arabidopsis thaliana. It enables optical control of protein-protein interactions and has been used to regulate processes including transcription, protein localization, protein secretion, and, when coupled to BAX, light-triggered apoptosis.
Need inducible protein relocalization or recruitment
DerivedCry2/CIB is a genetically encoded blue-light-activated protein dimerization module derived from Arabidopsis thaliana. It enables optical control of protein-protein interactions and has been used to regulate processes including transcription, protein localization, protein secretion, and, when coupled to BAX, light-triggered apoptosis.
Need precise spatiotemporal control with light input
DerivedCry2/CIB is a genetically encoded blue-light-activated protein dimerization module derived from Arabidopsis thaliana. It enables optical control of protein-protein interactions and has been used to regulate processes including transcription, protein localization, protein secretion, and, when coupled to BAX, light-triggered apoptosis.
Need tighter control over gene expression timing or amplitude
DerivedCry2/CIB is a genetically encoded blue-light-activated protein dimerization module derived from Arabidopsis thaliana. It enables optical control of protein-protein interactions and has been used to regulate processes including transcription, protein localization, protein secretion, and, when coupled to BAX, light-triggered apoptosis.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
HeterodimerizationHeterodimerizationHeterodimerizationlight-gated protein-protein interactionlight-gated protein-protein interactionTechniques
No technique tags yet.
Target processes
localizationrecombinationtranscriptionInput: 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: reporterswitch architecture: multi componentswitch architecture: recruitment
Reported experimental design considerations for optical dimerizer use include selecting the dimerizer system, photoexcitation sources, and coordinated imaging reporters. The evidence supports implementation by domain fusion to effectors such as BAX, but it does not provide construct architecture, cofactor requirements, or delivery details.
The supplied evidence does not report quantitative performance metrics such as activation wavelength range, kinetics, dynamic range, or reversibility. The apoptosis-focused evidence also indicates that light-independent cell death can occur and required optimization of photophysical properties to improve control.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Source 2primary paper2018The Scholarship East Carolina University's Institutional Repository (East Carolina University)
Ranked Claims
The reported optimization efforts aimed to reduce light-independent cell death and improve experimental control by manipulating photophysical properties of the Cry2/CIB interaction.
We also report results of experimental efforts to optimize our optogenetic switch to reduce light-independent cell death (dark activation), and to enhance experimental control of our switch by manipulating photophysical properties associated with the Cry2/CIB interaction.
The Cry2/CIB module used in conjunction with BAX enabled light-mediated initiation of mitochondrial outer membrane permeabilization and downstream apoptosis.
Previously, we have employed Cryptochrome 2 (Cry2)/CIB, a blue light photoreceptor protein - protein dimerization module from A. thaliana in conjunction with BAX, an OMM targeting pro-apoptotic protein, for light-mediated initiation of mitochondrial outer membrane permeabilization (MOMP) and downstream apoptosis.
CRY2/CIB and UVR8/UVR8 are presented for controlling transcription, protein localization, and protein secretion using light.
We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8, for use in controlling transcription, protein localization, and protein secretion using light.
CRY2/CIB and UVR8/UVR8 are presented for controlling transcription, protein localization, and protein secretion using light.
We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8, for use in controlling transcription, protein localization, and protein secretion using light.
CRY2/CIB and UVR8/UVR8 are presented for controlling transcription, protein localization, and protein secretion using light.
We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8, for use in controlling transcription, protein localization, and protein secretion using light.
CRY2/CIB and UVR8/UVR8 are presented for controlling transcription, protein localization, and protein secretion using light.
We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8, for use in controlling transcription, protein localization, and protein secretion using light.
CRY2/CIB and UVR8/UVR8 are presented for controlling transcription, protein localization, and protein secretion using light.
We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8, for use in controlling transcription, protein localization, and protein secretion using light.
CRY2/CIB and UVR8/UVR8 are presented for controlling transcription, protein localization, and protein secretion using light.
We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8, for use in controlling transcription, protein localization, and protein secretion using light.
CRY2/CIB and UVR8/UVR8 are presented for controlling transcription, protein localization, and protein secretion using light.
We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8, for use in controlling transcription, protein localization, and protein secretion using light.
CRY2/CIB and UVR8/UVR8 are presented for controlling transcription, protein localization, and protein secretion using light.
We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8, for use in controlling transcription, protein localization, and protein secretion using light.
CRY2/CIB and UVR8/UVR8 are presented for controlling transcription, protein localization, and protein secretion using light.
We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8, for use in controlling transcription, protein localization, and protein secretion using light.
CRY2/CIB and UVR8/UVR8 are presented for controlling transcription, protein localization, and protein secretion using light.
We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8, for use in controlling transcription, protein localization, and protein secretion using light.
CRY2/CIB and UVR8/UVR8 are presented for controlling transcription, protein localization, and protein secretion using light.
We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8, for use in controlling transcription, protein localization, and protein secretion using light.
Design of optical dimerizer experiments includes choosing a dimerizer system, photoexcitation sources, and coordinated imaging reporters.
Here we discuss the design of optical dimerizer experiments, including choice of a dimerizer system, photoexcitation sources, and the coordinate use of imaging reporters.
Design of optical dimerizer experiments includes choosing a dimerizer system, photoexcitation sources, and coordinated imaging reporters.
Here we discuss the design of optical dimerizer experiments, including choice of a dimerizer system, photoexcitation sources, and the coordinate use of imaging reporters.
Design of optical dimerizer experiments includes choosing a dimerizer system, photoexcitation sources, and coordinated imaging reporters.
Here we discuss the design of optical dimerizer experiments, including choice of a dimerizer system, photoexcitation sources, and the coordinate use of imaging reporters.
Design of optical dimerizer experiments includes choosing a dimerizer system, photoexcitation sources, and coordinated imaging reporters.
Here we discuss the design of optical dimerizer experiments, including choice of a dimerizer system, photoexcitation sources, and the coordinate use of imaging reporters.
Design of optical dimerizer experiments includes choosing a dimerizer system, photoexcitation sources, and coordinated imaging reporters.
Here we discuss the design of optical dimerizer experiments, including choice of a dimerizer system, photoexcitation sources, and the coordinate use of imaging reporters.
Design of optical dimerizer experiments includes choosing a dimerizer system, photoexcitation sources, and coordinated imaging reporters.
Here we discuss the design of optical dimerizer experiments, including choice of a dimerizer system, photoexcitation sources, and the coordinate use of imaging reporters.
Design of optical dimerizer experiments includes choosing a dimerizer system, photoexcitation sources, and coordinated imaging reporters.
Here we discuss the design of optical dimerizer experiments, including choice of a dimerizer system, photoexcitation sources, and the coordinate use of imaging reporters.
Design of optical dimerizer experiments includes choosing a dimerizer system, photoexcitation sources, and coordinated imaging reporters.
Here we discuss the design of optical dimerizer experiments, including choice of a dimerizer system, photoexcitation sources, and the coordinate use of imaging reporters.
Design of optical dimerizer experiments includes choosing a dimerizer system, photoexcitation sources, and coordinated imaging reporters.
Here we discuss the design of optical dimerizer experiments, including choice of a dimerizer system, photoexcitation sources, and the coordinate use of imaging reporters.
Design of optical dimerizer experiments includes choosing a dimerizer system, photoexcitation sources, and coordinated imaging reporters.
Here we discuss the design of optical dimerizer experiments, including choice of a dimerizer system, photoexcitation sources, and the coordinate use of imaging reporters.
Numerous new and versatile optical dimerizer systems have been developed in recent years.
In recent years, numerous new and versatile dimerizer systems have been developed.
Numerous new and versatile optical dimerizer systems have been developed in recent years.
In recent years, numerous new and versatile dimerizer systems have been developed.
Numerous new and versatile optical dimerizer systems have been developed in recent years.
In recent years, numerous new and versatile dimerizer systems have been developed.
Numerous new and versatile optical dimerizer systems have been developed in recent years.
In recent years, numerous new and versatile dimerizer systems have been developed.
Numerous new and versatile optical dimerizer systems have been developed in recent years.
In recent years, numerous new and versatile dimerizer systems have been developed.
Numerous new and versatile optical dimerizer systems have been developed in recent years.
In recent years, numerous new and versatile dimerizer systems have been developed.
Numerous new and versatile optical dimerizer systems have been developed in recent years.
In recent years, numerous new and versatile dimerizer systems have been developed.
Numerous new and versatile optical dimerizer systems have been developed in recent years.
In recent years, numerous new and versatile dimerizer systems have been developed.
Numerous new and versatile optical dimerizer systems have been developed in recent years.
In recent years, numerous new and versatile dimerizer systems have been developed.
Numerous new and versatile optical dimerizer systems have been developed in recent years.
In recent years, numerous new and versatile dimerizer systems have been developed.
Optical dimerizers are genetically encoded actuators that enable light control of protein-protein interactions.
Genetically encoded actuators that allow control of protein-protein interactions using light, termed 'optical dimerizers'
Optical dimerizers are genetically encoded actuators that enable light control of protein-protein interactions.
Genetically encoded actuators that allow control of protein-protein interactions using light, termed 'optical dimerizers'
Optical dimerizers are genetically encoded actuators that enable light control of protein-protein interactions.
Genetically encoded actuators that allow control of protein-protein interactions using light, termed 'optical dimerizers'
Optical dimerizers are genetically encoded actuators that enable light control of protein-protein interactions.
Genetically encoded actuators that allow control of protein-protein interactions using light, termed 'optical dimerizers'
Optical dimerizers are genetically encoded actuators that enable light control of protein-protein interactions.
Genetically encoded actuators that allow control of protein-protein interactions using light, termed 'optical dimerizers'
Optical dimerizers are genetically encoded actuators that enable light control of protein-protein interactions.
Genetically encoded actuators that allow control of protein-protein interactions using light, termed 'optical dimerizers'
Optical dimerizers are genetically encoded actuators that enable light control of protein-protein interactions.
Genetically encoded actuators that allow control of protein-protein interactions using light, termed 'optical dimerizers'
Optical dimerizers are genetically encoded actuators that enable light control of protein-protein interactions.
Genetically encoded actuators that allow control of protein-protein interactions using light, termed 'optical dimerizers'
Optical dimerizers are genetically encoded actuators that enable light control of protein-protein interactions.
Genetically encoded actuators that allow control of protein-protein interactions using light, termed 'optical dimerizers'
Optical dimerizers are genetically encoded actuators that enable light control of protein-protein interactions.
Genetically encoded actuators that allow control of protein-protein interactions using light, termed 'optical dimerizers'
A pulse-controlled LED device is provided for experiments requiring extended light treatments.
we provide instructions and software for constructing a pulse-controlled LED device for use in experiments requiring extended light treatments
A pulse-controlled LED device is provided for experiments requiring extended light treatments.
we provide instructions and software for constructing a pulse-controlled LED device for use in experiments requiring extended light treatments
A pulse-controlled LED device is provided for experiments requiring extended light treatments.
we provide instructions and software for constructing a pulse-controlled LED device for use in experiments requiring extended light treatments
A pulse-controlled LED device is provided for experiments requiring extended light treatments.
we provide instructions and software for constructing a pulse-controlled LED device for use in experiments requiring extended light treatments
A pulse-controlled LED device is provided for experiments requiring extended light treatments.
we provide instructions and software for constructing a pulse-controlled LED device for use in experiments requiring extended light treatments
A pulse-controlled LED device is provided for experiments requiring extended light treatments.
we provide instructions and software for constructing a pulse-controlled LED device for use in experiments requiring extended light treatments
A pulse-controlled LED device is provided for experiments requiring extended light treatments.
we provide instructions and software for constructing a pulse-controlled LED device for use in experiments requiring extended light treatments
A pulse-controlled LED device is provided for experiments requiring extended light treatments.
we provide instructions and software for constructing a pulse-controlled LED device for use in experiments requiring extended light treatments
A pulse-controlled LED device is provided for experiments requiring extended light treatments.
we provide instructions and software for constructing a pulse-controlled LED device for use in experiments requiring extended light treatments
A pulse-controlled LED device is provided for experiments requiring extended light treatments.
we provide instructions and software for constructing a pulse-controlled LED device for use in experiments requiring extended light treatments
Approval Evidence
3 sources3 linked approval claimsfirst-pass slug cry2-cib
Previous work has employed the Cryptochrome 2 (Cry2)/CIB, a blue light activated protein – protein dimerization module from A. thaliana
Source:
Previously, we have employed Cryptochrome 2 (Cry2)/CIB, a blue light photoreceptor protein - protein dimerization module from A. thaliana in conjunction with BAX
Source:
We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8
Source:
optimization goalsupports
The reported optimization efforts aimed to reduce light-independent cell death and improve experimental control by manipulating photophysical properties of the Cry2/CIB interaction.
We also report results of experimental efforts to optimize our optogenetic switch to reduce light-independent cell death (dark activation), and to enhance experimental control of our switch by manipulating photophysical properties associated with the Cry2/CIB interaction.
Source:
tool functionsupports
The Cry2/CIB module used in conjunction with BAX enabled light-mediated initiation of mitochondrial outer membrane permeabilization and downstream apoptosis.
Previously, we have employed Cryptochrome 2 (Cry2)/CIB, a blue light photoreceptor protein - protein dimerization module from A. thaliana in conjunction with BAX, an OMM targeting pro-apoptotic protein, for light-mediated initiation of mitochondrial outer membrane permeabilization (MOMP) and downstream apoptosis.
Source:
application scopesupports
CRY2/CIB and UVR8/UVR8 are presented for controlling transcription, protein localization, and protein secretion using light.
We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8, for use in controlling transcription, protein localization, and protein secretion using light.
Source:
Comparisons
Source-backed strengths
The system is explicitly described as a blue-light-activated protein-protein dimerization module from A. thaliana and has been used across multiple application classes, including transcriptional control, protein localization, protein secretion, and BAX-mediated apoptosis. In conjunction with BAX, it enabled light-mediated initiation of mitochondrial outer membrane permeabilization and downstream apoptosis.
Source:
We also report results of experimental efforts to optimize our optogenetic switch to reduce light-independent cell death (dark activation), and to enhance experimental control of our switch by manipulating photophysical properties associated with the Cry2/CIB interaction.
Compared with iLID/SspB
Cry2/CIB and iLID/SspB address a similar problem space because they share localization, recombination, transcription.
Shared frame: same top-level item type; shared target processes: localization, recombination, transcription; shared mechanisms: heterodimerization; same primary input modality: light
Relative tradeoffs: appears more independently replicated.
Cry2/CIB and single-component optogenetic tools for inducible RhoA GTPase signaling address a similar problem space because they share localization, recombination, transcription.
Shared frame: same top-level item type; shared target processes: localization, recombination, transcription; shared mechanisms: heterodimerization; same primary input modality: light
Strengths here: appears more independently replicated; looks easier to implement in practice.
Compared with UVR8/UVR8
Cry2/CIB and UVR8/UVR8 address a similar problem space because they share localization, recombination, transcription.
Shared frame: same top-level item type; shared target processes: localization, recombination, transcription; shared mechanisms: heterodimerization; same primary input modality: light
Strengths here: appears more independently replicated; looks easier to implement in practice.
Ranked Citations
- 1.