Fabrication of duplex DNA microarrays incorporating methyl-5-cytosine.
Identifieur interne : 002255 ( Main/Curation ); précédent : 002254; suivant : 002256Fabrication of duplex DNA microarrays incorporating methyl-5-cytosine.
Auteurs : Christopher L. Warren [États-Unis] ; Jianfei Zhao ; Kimberly Glass ; Vikas Rishi ; Aseem Z. Ansari ; Charles VinsonSource :
- Lab on a chip [ 1473-0189 ] ; 2012.
Descripteurs français
- KwdFr :
- 5-Méthyl-cytosine (immunologie), 5-Méthyl-cytosine (métabolisme), ADN (métabolisme), Anticorps (immunologie), Ilots CpG, Méthylation de l'ADN, Protéine de liaison à l'élément de réponse à l'AMP cyclique (génétique), Séquençage par oligonucléotides en batterie (instrumentation), Test de retard de migration électrophorétique (instrumentation).
- MESH :
- génétique : Protéine de liaison à l'élément de réponse à l'AMP cyclique.
- immunologie : 5-Méthyl-cytosine, Anticorps.
- métabolisme : 5-Méthyl-cytosine, ADN, Séquençage par oligonucléotides en batterie, Test de retard de migration électrophorétique.
- Ilots CpG, Méthylation de l'ADN.
English descriptors
- KwdEn :
- 5-Methylcytosine (immunology), 5-Methylcytosine (metabolism), Antibodies (immunology), CpG Islands, Cyclic AMP Response Element-Binding Protein (genetics), DNA (metabolism), DNA Methylation, Electrophoretic Mobility Shift Assay (instrumentation), Oligonucleotide Array Sequence Analysis (instrumentation).
- MESH :
- chemical , genetics : Cyclic AMP Response Element-Binding Protein.
- chemical , immunology : 5-Methylcytosine, Antibodies.
- chemical , metabolism : 5-Methylcytosine, DNA.
- instrumentation : Electrophoretic Mobility Shift Assay, Oligonucleotide Array Sequence Analysis.
- CpG Islands, DNA Methylation.
Abstract
We synthesized customized double-stranded DNA microarrays including methyl-5-cytosine at CpG dinucleotides and produced all 163,555 possible 8-mers (un-, hemi-, and di-methylated) to gain insight into how methylation affects transcription factor binding. An antibody to methyl-5-cytidine showed greater binding to the methylated DNA, demonstrating efficient incorporation of methyl-5-cytosine into the synthesized DNA. In contrast, binding of the transcription factor CREB was inhibited by CpG methylation. This platform represents a powerful new technology to evaluate the effect of DNA methylation on protein binding in any sequence context.
DOI: 10.1039/c1lc20698b
PubMed: 22139143
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pubmed:22139143Le document en format XML
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<term>5-Methylcytosine (metabolism)</term>
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<term>CpG Islands</term>
<term>Cyclic AMP Response Element-Binding Protein (genetics)</term>
<term>DNA (metabolism)</term>
<term>DNA Methylation</term>
<term>Electrophoretic Mobility Shift Assay (instrumentation)</term>
<term>Oligonucleotide Array Sequence Analysis (instrumentation)</term>
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<keywords scheme="KwdFr" xml:lang="fr"><term>5-Méthyl-cytosine (immunologie)</term>
<term>5-Méthyl-cytosine (métabolisme)</term>
<term>ADN (métabolisme)</term>
<term>Anticorps (immunologie)</term>
<term>Ilots CpG</term>
<term>Méthylation de l'ADN</term>
<term>Protéine de liaison à l'élément de réponse à l'AMP cyclique (génétique)</term>
<term>Séquençage par oligonucléotides en batterie (instrumentation)</term>
<term>Test de retard de migration électrophorétique (instrumentation)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Cyclic AMP Response Element-Binding Protein</term>
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<term>Antibodies</term>
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<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>5-Methylcytosine</term>
<term>DNA</term>
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<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéine de liaison à l'élément de réponse à l'AMP cyclique</term>
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<keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>5-Méthyl-cytosine</term>
<term>Anticorps</term>
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<keywords scheme="MESH" qualifier="instrumentation" xml:lang="en"><term>Electrophoretic Mobility Shift Assay</term>
<term>Oligonucleotide Array Sequence Analysis</term>
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<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>5-Méthyl-cytosine</term>
<term>ADN</term>
<term>Séquençage par oligonucléotides en batterie</term>
<term>Test de retard de migration électrophorétique</term>
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<keywords scheme="MESH" xml:lang="en"><term>CpG Islands</term>
<term>DNA Methylation</term>
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<keywords scheme="MESH" xml:lang="fr"><term>Ilots CpG</term>
<term>Méthylation de l'ADN</term>
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<front><div type="abstract" xml:lang="en">We synthesized customized double-stranded DNA microarrays including methyl-5-cytosine at CpG dinucleotides and produced all 163,555 possible 8-mers (un-, hemi-, and di-methylated) to gain insight into how methylation affects transcription factor binding. An antibody to methyl-5-cytidine showed greater binding to the methylated DNA, demonstrating efficient incorporation of methyl-5-cytosine into the synthesized DNA. In contrast, binding of the transcription factor CREB was inhibited by CpG methylation. This platform represents a powerful new technology to evaluate the effect of DNA methylation on protein binding in any sequence context.</div>
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