Source 1primary paper2017Scientific Reports
Toolkit/CRY2-TET1-CD fusion
CRY2-TET1-CD fusion
Also known as: CRY2 linked to TET1-CD
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
CRY2-TET1-CD fusion is an optogenetic epigenome-editing component in which the Arabidopsis CRY2 photoreceptor is fused to the TET1 catalytic domain and used with a CIB1-TALE targeting partner. In the reported system, blue-light illumination induced co-localization at the Ascl1 promoter and selectively altered local methylation state with associated regulation of gene activity.
Usefulness & Problems
Why this is useful
This tool enables light-gated, locus-specific recruitment of a DNA methylation-editing activity to a defined genomic site. In the reported study, it provided spatiotemporal control over epigenetic editing at the Ascl1 promoter in neural stem cells.
Source:
Herein, we have developed an optogenetic toolbox fused to the catalytic domain (CD) of DNA-methyltransferase3A (DNMT3A-CD) or Ten-Eleven Dioxygenase-1 (TET1-CD) for loci-specific alteration of the methylation state at the promoter of Ascl1 (Mash1). Optogenetical protein pairs, CRY2 linked to DNMT3A-CD or TET1-CD and CIB1 fused to a Transcription Activator-Like Element (TALE) locating an Ascl1 promoter region, were designed for site specific epigenetic editing.
Problem solved
It addresses the problem of how to control epigenetic editing at a specific promoter with temporal precision rather than constitutive targeting. The reported implementation uses CRY2/CIB1 optogenetic association and TALE-based DNA recognition to direct TET1 catalytic activity to the Ascl1 promoter.
Source:
Herein, we have developed an optogenetic toolbox fused to the catalytic domain (CD) of DNA-methyltransferase3A (DNMT3A-CD) or Ten-Eleven Dioxygenase-1 (TET1-CD) for loci-specific alteration of the methylation state at the promoter of Ascl1 (Mash1). Optogenetical protein pairs, CRY2 linked to DNMT3A-CD or TET1-CD and CIB1 fused to a Transcription Activator-Like Element (TALE) locating an Ascl1 promoter region, were designed for site specific epigenetic editing.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
epigenetic editing of dna methylation statelight-induced heterodimerization/co-localizationlocus-specific dna targeting by tale bindinglocus-specific recruitment via tale dna bindingTechniques
Computational DesignTarget processes
editinglocalizationtranscriptionInput: Light
Implementation Constraints
The reported design uses CRY2 fused to TET1-CD and CIB1 fused to a TALE that recognizes an Ascl1 promoter region. Function depends on optimized blue-light illumination and on co-expression of both fusion proteins for light-induced promoter co-localization.
The supplied evidence supports activity only at the Ascl1 promoter in the context of one 2017 study, so generality across loci, cell types, and organisms is not established here. Quantitative performance metrics, off-target effects, reversibility, and delivery constraints are not described in the provided evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
The spatiotemporal association of the fusion proteins selectively altered methylation state and regulated gene activity at the Ascl1 promoter.
We found that this spatiotemporal association of the fusion proteins selectively alters the methylation state and also regulates gene activity.
The spatiotemporal association of the fusion proteins selectively altered methylation state and regulated gene activity at the Ascl1 promoter.
We found that this spatiotemporal association of the fusion proteins selectively alters the methylation state and also regulates gene activity.
The spatiotemporal association of the fusion proteins selectively altered methylation state and regulated gene activity at the Ascl1 promoter.
We found that this spatiotemporal association of the fusion proteins selectively alters the methylation state and also regulates gene activity.
The spatiotemporal association of the fusion proteins selectively altered methylation state and regulated gene activity at the Ascl1 promoter.
We found that this spatiotemporal association of the fusion proteins selectively alters the methylation state and also regulates gene activity.
The spatiotemporal association of the fusion proteins selectively altered methylation state and regulated gene activity at the Ascl1 promoter.
We found that this spatiotemporal association of the fusion proteins selectively alters the methylation state and also regulates gene activity.
The spatiotemporal association of the fusion proteins selectively altered methylation state and regulated gene activity at the Ascl1 promoter.
We found that this spatiotemporal association of the fusion proteins selectively alters the methylation state and also regulates gene activity.
The spatiotemporal association of the fusion proteins selectively altered methylation state and regulated gene activity at the Ascl1 promoter.
We found that this spatiotemporal association of the fusion proteins selectively alters the methylation state and also regulates gene activity.
Optimized blue-light illumination triggered co-localization of the TALE construct with DNMT3A-CD or TET1-CD fusion proteins at the targeted Ascl1 promoter site.
Optimized blue-light illumination triggered the co-localization of TALE constructs with DNMT3A-CD or TET1-CD fusion proteins at the targeted site of the Ascl1 promoter.
Optimized blue-light illumination triggered co-localization of the TALE construct with DNMT3A-CD or TET1-CD fusion proteins at the targeted Ascl1 promoter site.
Optimized blue-light illumination triggered the co-localization of TALE constructs with DNMT3A-CD or TET1-CD fusion proteins at the targeted site of the Ascl1 promoter.
Optimized blue-light illumination triggered co-localization of the TALE construct with DNMT3A-CD or TET1-CD fusion proteins at the targeted Ascl1 promoter site.
Optimized blue-light illumination triggered the co-localization of TALE constructs with DNMT3A-CD or TET1-CD fusion proteins at the targeted site of the Ascl1 promoter.
Optimized blue-light illumination triggered co-localization of the TALE construct with DNMT3A-CD or TET1-CD fusion proteins at the targeted Ascl1 promoter site.
Optimized blue-light illumination triggered the co-localization of TALE constructs with DNMT3A-CD or TET1-CD fusion proteins at the targeted site of the Ascl1 promoter.
Optimized blue-light illumination triggered co-localization of the TALE construct with DNMT3A-CD or TET1-CD fusion proteins at the targeted Ascl1 promoter site.
Optimized blue-light illumination triggered the co-localization of TALE constructs with DNMT3A-CD or TET1-CD fusion proteins at the targeted site of the Ascl1 promoter.
Optimized blue-light illumination triggered co-localization of the TALE construct with DNMT3A-CD or TET1-CD fusion proteins at the targeted Ascl1 promoter site.
Optimized blue-light illumination triggered the co-localization of TALE constructs with DNMT3A-CD or TET1-CD fusion proteins at the targeted site of the Ascl1 promoter.
Optimized blue-light illumination triggered co-localization of the TALE construct with DNMT3A-CD or TET1-CD fusion proteins at the targeted Ascl1 promoter site.
Optimized blue-light illumination triggered the co-localization of TALE constructs with DNMT3A-CD or TET1-CD fusion proteins at the targeted site of the Ascl1 promoter.
The paper reports development of an optogenetic toolbox using CRY2/CIB1 fusions with DNMT3A-CD or TET1-CD and a TALE to enable locus-specific epigenetic editing at the Ascl1 promoter.
Herein, we have developed an optogenetic toolbox fused to the catalytic domain (CD) of DNA-methyltransferase3A (DNMT3A-CD) or Ten-Eleven Dioxygenase-1 (TET1-CD) for loci-specific alteration of the methylation state at the promoter of Ascl1 (Mash1). Optogenetical protein pairs, CRY2 linked to DNMT3A-CD or TET1-CD and CIB1 fused to a Transcription Activator-Like Element (TALE) locating an Ascl1 promoter region, were designed for site specific epigenetic editing.
The paper reports development of an optogenetic toolbox using CRY2/CIB1 fusions with DNMT3A-CD or TET1-CD and a TALE to enable locus-specific epigenetic editing at the Ascl1 promoter.
Herein, we have developed an optogenetic toolbox fused to the catalytic domain (CD) of DNA-methyltransferase3A (DNMT3A-CD) or Ten-Eleven Dioxygenase-1 (TET1-CD) for loci-specific alteration of the methylation state at the promoter of Ascl1 (Mash1). Optogenetical protein pairs, CRY2 linked to DNMT3A-CD or TET1-CD and CIB1 fused to a Transcription Activator-Like Element (TALE) locating an Ascl1 promoter region, were designed for site specific epigenetic editing.
The paper reports development of an optogenetic toolbox using CRY2/CIB1 fusions with DNMT3A-CD or TET1-CD and a TALE to enable locus-specific epigenetic editing at the Ascl1 promoter.
Herein, we have developed an optogenetic toolbox fused to the catalytic domain (CD) of DNA-methyltransferase3A (DNMT3A-CD) or Ten-Eleven Dioxygenase-1 (TET1-CD) for loci-specific alteration of the methylation state at the promoter of Ascl1 (Mash1). Optogenetical protein pairs, CRY2 linked to DNMT3A-CD or TET1-CD and CIB1 fused to a Transcription Activator-Like Element (TALE) locating an Ascl1 promoter region, were designed for site specific epigenetic editing.
The paper reports development of an optogenetic toolbox using CRY2/CIB1 fusions with DNMT3A-CD or TET1-CD and a TALE to enable locus-specific epigenetic editing at the Ascl1 promoter.
Herein, we have developed an optogenetic toolbox fused to the catalytic domain (CD) of DNA-methyltransferase3A (DNMT3A-CD) or Ten-Eleven Dioxygenase-1 (TET1-CD) for loci-specific alteration of the methylation state at the promoter of Ascl1 (Mash1). Optogenetical protein pairs, CRY2 linked to DNMT3A-CD or TET1-CD and CIB1 fused to a Transcription Activator-Like Element (TALE) locating an Ascl1 promoter region, were designed for site specific epigenetic editing.
The paper reports development of an optogenetic toolbox using CRY2/CIB1 fusions with DNMT3A-CD or TET1-CD and a TALE to enable locus-specific epigenetic editing at the Ascl1 promoter.
Herein, we have developed an optogenetic toolbox fused to the catalytic domain (CD) of DNA-methyltransferase3A (DNMT3A-CD) or Ten-Eleven Dioxygenase-1 (TET1-CD) for loci-specific alteration of the methylation state at the promoter of Ascl1 (Mash1). Optogenetical protein pairs, CRY2 linked to DNMT3A-CD or TET1-CD and CIB1 fused to a Transcription Activator-Like Element (TALE) locating an Ascl1 promoter region, were designed for site specific epigenetic editing.
The paper reports development of an optogenetic toolbox using CRY2/CIB1 fusions with DNMT3A-CD or TET1-CD and a TALE to enable locus-specific epigenetic editing at the Ascl1 promoter.
Herein, we have developed an optogenetic toolbox fused to the catalytic domain (CD) of DNA-methyltransferase3A (DNMT3A-CD) or Ten-Eleven Dioxygenase-1 (TET1-CD) for loci-specific alteration of the methylation state at the promoter of Ascl1 (Mash1). Optogenetical protein pairs, CRY2 linked to DNMT3A-CD or TET1-CD and CIB1 fused to a Transcription Activator-Like Element (TALE) locating an Ascl1 promoter region, were designed for site specific epigenetic editing.
The paper reports development of an optogenetic toolbox using CRY2/CIB1 fusions with DNMT3A-CD or TET1-CD and a TALE to enable locus-specific epigenetic editing at the Ascl1 promoter.
Herein, we have developed an optogenetic toolbox fused to the catalytic domain (CD) of DNA-methyltransferase3A (DNMT3A-CD) or Ten-Eleven Dioxygenase-1 (TET1-CD) for loci-specific alteration of the methylation state at the promoter of Ascl1 (Mash1). Optogenetical protein pairs, CRY2 linked to DNMT3A-CD or TET1-CD and CIB1 fused to a Transcription Activator-Like Element (TALE) locating an Ascl1 promoter region, were designed for site specific epigenetic editing.
Approval Evidence
1 source3 linked approval claimsfirst-pass slug cry2-tet1-cd-fusion
Optogenetical protein pairs, CRY2 linked to DNMT3A-CD or TET1-CD and CIB1 fused to a Transcription Activator-Like Element (TALE) locating an Ascl1 promoter region, were designed for site specific epigenetic editing.
Source:
epigenetic editing effectsupports
The spatiotemporal association of the fusion proteins selectively altered methylation state and regulated gene activity at the Ascl1 promoter.
We found that this spatiotemporal association of the fusion proteins selectively alters the methylation state and also regulates gene activity.
Source:
light induced colocalizationsupports
Optimized blue-light illumination triggered co-localization of the TALE construct with DNMT3A-CD or TET1-CD fusion proteins at the targeted Ascl1 promoter site.
Optimized blue-light illumination triggered the co-localization of TALE constructs with DNMT3A-CD or TET1-CD fusion proteins at the targeted site of the Ascl1 promoter.
Source:
tool developmentsupports
The paper reports development of an optogenetic toolbox using CRY2/CIB1 fusions with DNMT3A-CD or TET1-CD and a TALE to enable locus-specific epigenetic editing at the Ascl1 promoter.
Herein, we have developed an optogenetic toolbox fused to the catalytic domain (CD) of DNA-methyltransferase3A (DNMT3A-CD) or Ten-Eleven Dioxygenase-1 (TET1-CD) for loci-specific alteration of the methylation state at the promoter of Ascl1 (Mash1). Optogenetical protein pairs, CRY2 linked to DNMT3A-CD or TET1-CD and CIB1 fused to a Transcription Activator-Like Element (TALE) locating an Ascl1 promoter region, were designed for site specific epigenetic editing.
Source:
Comparisons
Source-backed strengths
The system was specifically developed for site-specific epigenetic editing using modular CRY2/CIB1 and TALE fusions. Under optimized blue-light illumination, the TALE construct co-localized with the TET1-CD fusion at the targeted Ascl1 promoter, and this association selectively altered methylation state and regulated gene activity.
Ranked Citations
- 1.