Source 1primary paper2017Scientific Reports
Toolkit/CRY2-DNMT3A-CD fusion
CRY2-DNMT3A-CD fusion
Also known as: CRY2 linked to DNMT3A-CD
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
CRY2-DNMT3A-CD fusion is an optogenetic epigenetic editing component in which the photoreceptor CRY2 is fused to the catalytic domain of DNMT3A and used with a CIB1-TALE targeting partner. In the reported system, blue-light stimulation induced recruitment of DNMT3A-CD to the Ascl1 promoter, enabling locus-specific methylation changes and regulation of gene activity.
Usefulness & Problems
Why this is useful
This tool provides light-gated control over locus-specific DNA methylation editing at a defined genomic promoter. It is useful for studying how spatiotemporally controlled recruitment of a DNA methyltransferase catalytic domain affects promoter methylation state and transcriptional regulation.
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 selectively manipulating epigenetic state at a specific endogenous locus rather than globally altering DNA methylation. In the reported application, it enabled targeted editing of the Ascl1 promoter in neural stem cells through blue-light-dependent assembly of the editing complex.
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
light-induced heterodimerization/associationsite-specific epigenetic editing via dna methylationtargeted subcellular colocalization at a genomic locusTechniques
Computational DesignTarget processes
editinglocalizationtranscriptionInput: Light
Implementation Constraints
The reported implementation used a multi-component design consisting of CRY2 fused to DNMT3A-CD and CIB1 fused to a TALE that locates the Ascl1 promoter region. Blue-light illumination was used to trigger co-localization of these components at the target site; no additional construct architecture, delivery method, or cofactor requirements are specified in the supplied evidence.
The supplied evidence is limited to a single 2017 study focused on the Ascl1 promoter, so generalizability to other loci, cell types, or in vivo settings is not established here. Quantitative performance metrics, off-target methylation assessment, reversibility, and comparative benchmarking are not provided in the supplied 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-dnmt3a-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 reported to achieve optimized blue-light-triggered co-localization of the TALE construct with the CRY2-DNMT3A-CD fusion at the targeted Ascl1 promoter site. This spatiotemporal association selectively altered methylation state and regulated gene activity at that promoter, supporting functional locus-specific epigenetic editing.
Ranked Citations
- 1.