Source 1primary paper2017Scientific Reports
Toolkit/optogenetic epigenetic editing toolbox for Ascl1 promoter targeting
optogenetic epigenetic editing toolbox for Ascl1 promoter targeting
Also known as: optogenetic-epigenetic constructs, optogenetic toolbox
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The optogenetic epigenetic editing toolbox for Ascl1 promoter targeting is a multi-component light-responsive system that combines CRY2/CIB1 fusions, a TALE DNA-targeting module, and either the DNMT3A catalytic domain or TET1 catalytic domain. Under optimized blue-light illumination, it co-localizes these effectors at the Ascl1 promoter to alter local DNA methylation state and regulate Ascl1 gene activity.
Usefulness & Problems
Why this is useful
This toolbox is useful for locus-specific and light-gated manipulation of epigenetic state at the Ascl1 promoter. It addresses the need for spatiotemporal control over promoter methylation and associated transcriptional regulation using an optogenetic input.
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 helps solve the problem of selectively recruiting DNA methylation or demethylation activity to a defined endogenous promoter in a light-dependent manner. The reported application is targeted editing of the Ascl1 promoter methylation state to modulate gene activity.
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/co-localizationlocus-specific dna binding via tale targetingtargeted dna demethylationtargeted dna methylationTechniques
Computational DesignTarget processes
editinglocalizationtranscriptionInput: Light
Implementation Constraints
The construct architecture reported in the source uses CRY2/CIB1 light-responsive fusions together with a TALE targeted to the Ascl1 promoter and fused epigenetic effector catalytic domains from DNMT3A or TET1. Activation requires optimized blue-light illumination to trigger co-localization of the TALE construct with the DNMT3A-CD or TET1-CD fusion proteins at the target site.
The supplied evidence supports activity specifically at the Ascl1 promoter and does not establish performance at other loci or in other cell types beyond the reported neural stem cell context. Quantitative performance metrics, off-target effects, reversibility, and long-term stability 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 optogenetic-epigenetic-editing-toolbox-for-ascl1-promoter-targeting
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).
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 reported system enables selective alteration of methylation state and regulation of gene activity at the Ascl1 promoter. It supports recruitment of either DNMT3A-CD or TET1-CD through the same light-responsive framework, providing both targeted methylation and targeted demethylation modalities.
Ranked Citations
- 1.