Source 1primary paper2016Oncotarget
Toolkit/DNMT3A-CRY2/CIB1-TRF1 optogenetic telomere-targeting fusion construct system
DNMT3A-CRY2/CIB1-TRF1 optogenetic telomere-targeting fusion construct system
Also known as: CIB1-TRF1, DNMT3A-CRY2, optogenetic tools
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The DNMT3A-CRY2/CIB1-TRF1 system is a blue-light-responsive multi-component optogenetic construct that recruits human DNMT3A to telomeric regions through CRY2-CIB1 interaction and TRF1-mediated telomere binding. In the reported HeLa cell configuration, light activation selectively increased subtelomeric CpG methylation at six examined chromosome ends and was associated with progressive telomere lengthening over three cell generations.
Usefulness & Problems
Why this is useful
This system is useful for inducible, locus-directed epigenetic editing at chromosome ends, enabling optical control of subtelomeric DNA methylation rather than constitutive enzyme targeting. The reported modular architecture also supports substitution of other chromatin-modifying enzymes for site-specific recruitment.
Problem solved
It addresses the problem of selectively assembling a DNA methyltransferase at telomeric/subtelomeric regions with temporal control using light. The reported application specifically enabled testing whether targeted subtelomeric CpG methylation influences telomere length dynamics in HeLa cells.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Techniques
Computational DesignTarget processes
recombinationInput: Light
Implementation Constraints
The reported design comprises a DNMT3A-CRY2 fusion and a CIB1-TRF1 fusion, with blue-light excitation used to assemble the complex at telomeric regions. TRF1 provides telomere-associated localization, while CRY2 and CIB1 provide the light-dependent interaction module. The evidence supports use in vivo in HeLa cells, but does not specify construct stoichiometry, expression method, cofactor requirements, or illumination hardware details.
The supplied evidence is limited to a single cited study and a specific reported context, with validation described for six chromosome ends in HeLa cells. The evidence does not provide quantitative performance details such as recruitment kinetics, reversibility, off-target methylation, illumination parameters, or performance in other cell types or organisms. The association with telomere lengthening is reported, but the supplied text does not establish broader generality or mechanistic sufficiency beyond that study.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Blue-light activation of the optogenetic telomere-targeting construct selectively increased subtelomeric CpG methylation at six examined chromosome ends.
Increased methylation was achieved selectively at subtelomeric CpG sites on the six examined chromosome ends specifically after blue-light activation
Section: abstract
examined chromosome ends 6
Blue-light activation of the optogenetic telomere-targeting construct selectively increased subtelomeric CpG methylation at six examined chromosome ends.
Increased methylation was achieved selectively at subtelomeric CpG sites on the six examined chromosome ends specifically after blue-light activation
Section: abstract
examined chromosome ends 6
Blue-light activation of the optogenetic telomere-targeting construct selectively increased subtelomeric CpG methylation at six examined chromosome ends.
Increased methylation was achieved selectively at subtelomeric CpG sites on the six examined chromosome ends specifically after blue-light activation
Section: abstract
examined chromosome ends 6
Blue-light activation of the optogenetic telomere-targeting construct selectively increased subtelomeric CpG methylation at six examined chromosome ends.
Increased methylation was achieved selectively at subtelomeric CpG sites on the six examined chromosome ends specifically after blue-light activation
Section: abstract
examined chromosome ends 6
Blue-light activation of the optogenetic telomere-targeting construct selectively increased subtelomeric CpG methylation at six examined chromosome ends.
Increased methylation was achieved selectively at subtelomeric CpG sites on the six examined chromosome ends specifically after blue-light activation
Section: abstract
examined chromosome ends 6
Blue-light activation of the optogenetic telomere-targeting construct selectively increased subtelomeric CpG methylation at six examined chromosome ends.
Increased methylation was achieved selectively at subtelomeric CpG sites on the six examined chromosome ends specifically after blue-light activation
Section: abstract
examined chromosome ends 6
Blue-light activation of the optogenetic telomere-targeting construct selectively increased subtelomeric CpG methylation at six examined chromosome ends.
Increased methylation was achieved selectively at subtelomeric CpG sites on the six examined chromosome ends specifically after blue-light activation
Section: abstract
examined chromosome ends 6
The fusion construct design is modular and allows substitution of other chromatin-modifying enzymes for locus-specific targeting.
The modular design of the fusion constructs presented here allows for the selective substitution of other chromatin modifying enzymes and for loci-specific targeting to regulate the epigenetic pathways at telomeres and other selected genomic regions of interest.
Section: abstract
The fusion construct design is modular and allows substitution of other chromatin-modifying enzymes for locus-specific targeting.
The modular design of the fusion constructs presented here allows for the selective substitution of other chromatin modifying enzymes and for loci-specific targeting to regulate the epigenetic pathways at telomeres and other selected genomic regions of interest.
Section: abstract
The fusion construct design is modular and allows substitution of other chromatin-modifying enzymes for locus-specific targeting.
The modular design of the fusion constructs presented here allows for the selective substitution of other chromatin modifying enzymes and for loci-specific targeting to regulate the epigenetic pathways at telomeres and other selected genomic regions of interest.
Section: abstract
The fusion construct design is modular and allows substitution of other chromatin-modifying enzymes for locus-specific targeting.
The modular design of the fusion constructs presented here allows for the selective substitution of other chromatin modifying enzymes and for loci-specific targeting to regulate the epigenetic pathways at telomeres and other selected genomic regions of interest.
Section: abstract
The fusion construct design is modular and allows substitution of other chromatin-modifying enzymes for locus-specific targeting.
The modular design of the fusion constructs presented here allows for the selective substitution of other chromatin modifying enzymes and for loci-specific targeting to regulate the epigenetic pathways at telomeres and other selected genomic regions of interest.
Section: abstract
The fusion construct design is modular and allows substitution of other chromatin-modifying enzymes for locus-specific targeting.
The modular design of the fusion constructs presented here allows for the selective substitution of other chromatin modifying enzymes and for loci-specific targeting to regulate the epigenetic pathways at telomeres and other selected genomic regions of interest.
Section: abstract
The fusion construct design is modular and allows substitution of other chromatin-modifying enzymes for locus-specific targeting.
The modular design of the fusion constructs presented here allows for the selective substitution of other chromatin modifying enzymes and for loci-specific targeting to regulate the epigenetic pathways at telomeres and other selected genomic regions of interest.
Section: abstract
Selective subtelomeric methylation induced by blue-light activation was associated with a progressive increase in telomere length over three generations of HeLa cell replications.
which resulted in progressive increase in telomere length over three generations of HeLa cell replications
Section: abstract
replication generations 3
Selective subtelomeric methylation induced by blue-light activation was associated with a progressive increase in telomere length over three generations of HeLa cell replications.
which resulted in progressive increase in telomere length over three generations of HeLa cell replications
Section: abstract
replication generations 3
Selective subtelomeric methylation induced by blue-light activation was associated with a progressive increase in telomere length over three generations of HeLa cell replications.
which resulted in progressive increase in telomere length over three generations of HeLa cell replications
Section: abstract
replication generations 3
Selective subtelomeric methylation induced by blue-light activation was associated with a progressive increase in telomere length over three generations of HeLa cell replications.
which resulted in progressive increase in telomere length over three generations of HeLa cell replications
Section: abstract
replication generations 3
Selective subtelomeric methylation induced by blue-light activation was associated with a progressive increase in telomere length over three generations of HeLa cell replications.
which resulted in progressive increase in telomere length over three generations of HeLa cell replications
Section: abstract
replication generations 3
Selective subtelomeric methylation induced by blue-light activation was associated with a progressive increase in telomere length over three generations of HeLa cell replications.
which resulted in progressive increase in telomere length over three generations of HeLa cell replications
Section: abstract
replication generations 3
Selective subtelomeric methylation induced by blue-light activation was associated with a progressive increase in telomere length over three generations of HeLa cell replications.
which resulted in progressive increase in telomere length over three generations of HeLa cell replications
Section: abstract
replication generations 3
A blue-light-responsive DNMT3A-CRY2/CIB1-TRF1 fusion system localized DNMT3A-CRY2 to telomeric regions.
CIB1 was fused to the telomere-associated protein telomere repeat binding factor-1 (TRF1), which localized the protein complex DNMT3A-CRY2 at telomeric regions upon excitation by blue-light monitored by single-molecule fluorescence analyses.
Section: abstract
A blue-light-responsive DNMT3A-CRY2/CIB1-TRF1 fusion system localized DNMT3A-CRY2 to telomeric regions.
CIB1 was fused to the telomere-associated protein telomere repeat binding factor-1 (TRF1), which localized the protein complex DNMT3A-CRY2 at telomeric regions upon excitation by blue-light monitored by single-molecule fluorescence analyses.
Section: abstract
A blue-light-responsive DNMT3A-CRY2/CIB1-TRF1 fusion system localized DNMT3A-CRY2 to telomeric regions.
CIB1 was fused to the telomere-associated protein telomere repeat binding factor-1 (TRF1), which localized the protein complex DNMT3A-CRY2 at telomeric regions upon excitation by blue-light monitored by single-molecule fluorescence analyses.
Section: abstract
A blue-light-responsive DNMT3A-CRY2/CIB1-TRF1 fusion system localized DNMT3A-CRY2 to telomeric regions.
CIB1 was fused to the telomere-associated protein telomere repeat binding factor-1 (TRF1), which localized the protein complex DNMT3A-CRY2 at telomeric regions upon excitation by blue-light monitored by single-molecule fluorescence analyses.
Section: abstract
A blue-light-responsive DNMT3A-CRY2/CIB1-TRF1 fusion system localized DNMT3A-CRY2 to telomeric regions.
CIB1 was fused to the telomere-associated protein telomere repeat binding factor-1 (TRF1), which localized the protein complex DNMT3A-CRY2 at telomeric regions upon excitation by blue-light monitored by single-molecule fluorescence analyses.
Section: abstract
A blue-light-responsive DNMT3A-CRY2/CIB1-TRF1 fusion system localized DNMT3A-CRY2 to telomeric regions.
CIB1 was fused to the telomere-associated protein telomere repeat binding factor-1 (TRF1), which localized the protein complex DNMT3A-CRY2 at telomeric regions upon excitation by blue-light monitored by single-molecule fluorescence analyses.
Section: abstract
A blue-light-responsive DNMT3A-CRY2/CIB1-TRF1 fusion system localized DNMT3A-CRY2 to telomeric regions.
CIB1 was fused to the telomere-associated protein telomere repeat binding factor-1 (TRF1), which localized the protein complex DNMT3A-CRY2 at telomeric regions upon excitation by blue-light monitored by single-molecule fluorescence analyses.
Section: abstract
Approval Evidence
1 source4 linked approval claimsfirst-pass slug dnmt3a-cry2-cib1-trf1-optogenetic-telomere-targeting-fusion-construct-system
Human DNA methyltransferase3A (DNMT3A) were assembled selectively at chromosome ends by fusion to cryptochrome 2 protein (CRY2) and its interacting complement, the basic helix loop helix protein-1 (CIB1). CIB1 was fused to the telomere-associated protein telomere repeat binding factor-1 (TRF1), which localized the protein complex DNMT3A-CRY2 at telomeric regions upon excitation by blue-light.
Source:
epigenetic modificationsupports
Blue-light activation of the optogenetic telomere-targeting construct selectively increased subtelomeric CpG methylation at six examined chromosome ends.
Increased methylation was achieved selectively at subtelomeric CpG sites on the six examined chromosome ends specifically after blue-light activation
Source:
modularitysupports
The fusion construct design is modular and allows substitution of other chromatin-modifying enzymes for locus-specific targeting.
The modular design of the fusion constructs presented here allows for the selective substitution of other chromatin modifying enzymes and for loci-specific targeting to regulate the epigenetic pathways at telomeres and other selected genomic regions of interest.
Source:
phenotypic effectsupports
Selective subtelomeric methylation induced by blue-light activation was associated with a progressive increase in telomere length over three generations of HeLa cell replications.
which resulted in progressive increase in telomere length over three generations of HeLa cell replications
Source:
targeted localizationsupports
A blue-light-responsive DNMT3A-CRY2/CIB1-TRF1 fusion system localized DNMT3A-CRY2 to telomeric regions.
CIB1 was fused to the telomere-associated protein telomere repeat binding factor-1 (TRF1), which localized the protein complex DNMT3A-CRY2 at telomeric regions upon excitation by blue-light monitored by single-molecule fluorescence analyses.
Source:
Comparisons
Source-backed strengths
Evidence from the cited study indicates selective blue-light-induced increases in subtelomeric CpG methylation at six chromosome ends examined. The construct uses TRF1 to localize the CIB1 fusion to telomeric regions and CRY2-CIB1 light responsiveness to recruit DNMT3A on demand. The induced methylation state was associated with a progressive increase in telomere length over three generations of HeLa cell replications.
Ranked Citations
- 1.