Toolkit/CRY2 and LOV-fused degron light-responsive repression/degradation system

CRY2 and LOV-fused degron light-responsive repression/degradation system

Multi-Component Switch·Research·Since 2017

Also known as: CRY2 and a LOV-fused degron system

Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.

Summary

The CRY2 and LOV-fused degron light-responsive repression/degradation system is a multi-component optogenetic platform reported in mammalian cells that uses light to simultaneously block transcription and deplete protein levels. It is based on Arabidopsis cryptochrome 2 (CRY2), which interacts with CIB1 upon illumination, together with a LOV-fused degron configuration.

Usefulness & Problems

Why this is useful

This system is useful because it couples light control of transcriptional repression with light-driven protein depletion in a single framework, enabling bidirectional regulation of gene expression. The cited work states that such tools enable precise bidirectional control of gene expression across a variety of cells and model systems.

Source:

These tools will allow precise, bi-directional control of gene expression in a variety of cells and model systems.

Source:

a system using CRY2 and a LOV-fused degron to simultaneously block transcription and deplete protein levels with light

Source:

a system using CRY2 and CIB1 to induce protein expression with light through stimulation of transcription

Problem solved

This tool addresses the problem of coordinating rapid, light-triggered suppression of gene output at more than one regulatory layer. Specifically, the evidence supports a design intended to simultaneously block transcription and deplete protein levels with light.

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

degradationtranscription

Input: Light

Implementation Constraints

The system is multi-component and relies on CRY2 together with CIB1, since CRY2-CIB1 interaction upon illumination is explicitly supported by the source. The nuclear clearing phenotype depended on a dimerization domain in CRY2-fused transcriptional activators, indicating that construct architecture and domain fusion are important design variables; the evidence otherwise gives limited practical detail on expression, delivery, or cofactor requirements.

The supplied evidence does not provide quantitative performance metrics such as repression magnitude, degradation kinetics, reversibility, or wavelength dependence for the combined CRY2/LOV-degron configuration. Independent replication beyond the cited 2017 study is not provided, and validation details for organisms or cell types using the exact combined system are limited.

Validation

Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication

Supporting Sources

Ranked Claims

Claim 1capability statementsupports2017Source 1needs review

These tools enable precise bidirectional control of gene expression in a variety of cells and model systems.

These tools will allow precise, bi-directional control of gene expression in a variety of cells and model systems.
Claim 2capability statementsupports2017Source 1needs review

These tools enable precise bidirectional control of gene expression in a variety of cells and model systems.

These tools will allow precise, bi-directional control of gene expression in a variety of cells and model systems.
Claim 3capability statementsupports2017Source 1needs review

These tools enable precise bidirectional control of gene expression in a variety of cells and model systems.

These tools will allow precise, bi-directional control of gene expression in a variety of cells and model systems.
Claim 4capability statementsupports2017Source 1needs review

These tools enable precise bidirectional control of gene expression in a variety of cells and model systems.

These tools will allow precise, bi-directional control of gene expression in a variety of cells and model systems.
Claim 5capability statementsupports2017Source 1needs review

These tools enable precise bidirectional control of gene expression in a variety of cells and model systems.

These tools will allow precise, bi-directional control of gene expression in a variety of cells and model systems.
Claim 6capability statementsupports2017Source 1needs review

These tools enable precise bidirectional control of gene expression in a variety of cells and model systems.

These tools will allow precise, bi-directional control of gene expression in a variety of cells and model systems.
Claim 7capability statementsupports2017Source 1needs review

These tools enable precise bidirectional control of gene expression in a variety of cells and model systems.

These tools will allow precise, bi-directional control of gene expression in a variety of cells and model systems.
Claim 8interaction propertysupports2017Source 1needs review

CRY2 and CIB1 interact upon light illumination.

CRY2 and CIB1, Arabidopsis proteins that interact upon light illumination
Claim 9mechanistic dependencysupports2017Source 1needs review

The nuclear clearing phenotype depended on the presence of a dimerization domain in CRY2-fused transcriptional activators.

The nuclear clearing phenotype was dependent on the presence of a dimerization domain contained within the CRY2-fused transcriptional activators.
Claim 10phenotype observationsupports2017Source 1needs review

In mammalian cells, CRY2-tethered proteins showed light-dependent redistribution and clearing within the nucleus.

While adopting this approach to regulate transcription in mammalian cells, we observed light-dependent redistribution and clearing of CRY2-tethered proteins within the nucleus.
Claim 11tool functionsupports2017Source 1needs review

A CRY2-CIB1 system was developed to induce protein expression with light through stimulation of transcription.

a system using CRY2 and CIB1 to induce protein expression with light through stimulation of transcription
Claim 12tool functionsupports2017Source 1needs review

A CRY2-CIB1 system was developed to induce protein expression with light through stimulation of transcription.

a system using CRY2 and CIB1 to induce protein expression with light through stimulation of transcription
Claim 13tool functionsupports2017Source 1needs review

A CRY2-CIB1 system was developed to induce protein expression with light through stimulation of transcription.

a system using CRY2 and CIB1 to induce protein expression with light through stimulation of transcription
Claim 14tool functionsupports2017Source 1needs review

A CRY2-CIB1 system was developed to induce protein expression with light through stimulation of transcription.

a system using CRY2 and CIB1 to induce protein expression with light through stimulation of transcription
Claim 15tool functionsupports2017Source 1needs review

A CRY2-CIB1 system was developed to induce protein expression with light through stimulation of transcription.

a system using CRY2 and CIB1 to induce protein expression with light through stimulation of transcription
Claim 16tool functionsupports2017Source 1needs review

A CRY2-CIB1 system was developed to induce protein expression with light through stimulation of transcription.

a system using CRY2 and CIB1 to induce protein expression with light through stimulation of transcription
Claim 17tool functionsupports2017Source 1needs review

A CRY2-CIB1 system was developed to induce protein expression with light through stimulation of transcription.

a system using CRY2 and CIB1 to induce protein expression with light through stimulation of transcription
Claim 18tool functionsupports2017Source 1needs review

A CRY2 plus LOV-fused degron system was developed to simultaneously block transcription and deplete protein levels with light.

a system using CRY2 and a LOV-fused degron to simultaneously block transcription and deplete protein levels with light
Claim 19tool functionsupports2017Source 1needs review

A CRY2 plus LOV-fused degron system was developed to simultaneously block transcription and deplete protein levels with light.

a system using CRY2 and a LOV-fused degron to simultaneously block transcription and deplete protein levels with light
Claim 20tool functionsupports2017Source 1needs review

A CRY2 plus LOV-fused degron system was developed to simultaneously block transcription and deplete protein levels with light.

a system using CRY2 and a LOV-fused degron to simultaneously block transcription and deplete protein levels with light
Claim 21tool functionsupports2017Source 1needs review

A CRY2 plus LOV-fused degron system was developed to simultaneously block transcription and deplete protein levels with light.

a system using CRY2 and a LOV-fused degron to simultaneously block transcription and deplete protein levels with light
Claim 22tool functionsupports2017Source 1needs review

A CRY2 plus LOV-fused degron system was developed to simultaneously block transcription and deplete protein levels with light.

a system using CRY2 and a LOV-fused degron to simultaneously block transcription and deplete protein levels with light
Claim 23tool functionsupports2017Source 1needs review

A CRY2 plus LOV-fused degron system was developed to simultaneously block transcription and deplete protein levels with light.

a system using CRY2 and a LOV-fused degron to simultaneously block transcription and deplete protein levels with light
Claim 24tool functionsupports2017Source 1needs review

A CRY2 plus LOV-fused degron system was developed to simultaneously block transcription and deplete protein levels with light.

a system using CRY2 and a LOV-fused degron to simultaneously block transcription and deplete protein levels with light

Approval Evidence

1 source2 linked approval claimsfirst-pass slug cry2-and-lov-fused-degron-light-responsive-repression-degradation-system
a system using CRY2 and a LOV-fused degron to simultaneously block transcription and deplete protein levels with light

Source:

capability statementsupports

These tools enable precise bidirectional control of gene expression in a variety of cells and model systems.

These tools will allow precise, bi-directional control of gene expression in a variety of cells and model systems.

Source:

tool functionsupports

A CRY2 plus LOV-fused degron system was developed to simultaneously block transcription and deplete protein levels with light.

a system using CRY2 and a LOV-fused degron to simultaneously block transcription and deplete protein levels with light

Source:

Comparisons

Source-backed strengths

Evidence supports light-induced CRY2-CIB1 interaction and light-dependent redistribution/clearing of CRY2-tethered proteins within the nucleus in mammalian cells. The source also reports that these optogenetic tools provide precise bidirectional control of gene expression in multiple cellular and model-system contexts.

Ranked Citations

  1. 1.
    StructuralSource 1Nucleic Acids Research2017Claim 1Claim 2Claim 3

    Extracted from this source document.