Mapping Retinoic Acid-Dependant 5mC Derivatives in Mouse Embryonic Fibroblasts.
Methods Mol Biol
; 2019: 129-141, 2019.
Article
em En
| MEDLINE
| ID: mdl-31359394
ABSTRACT
Methylase-assisted bisulfite sequencing (MAB-seq) is a derivatization technique to evaluate the presence of 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC) at base-pair resolution. Although MAB-seq was originally designed to study these metabolites under steady-state conditions, we have developed an alternative protocol to evaluate the dynamics of 5-fC/5-caC accumulation in response to agonists, such as all-trans retinoic acid (ATRA). In addition, this protocol utilizes a lower quantity of the M.SssI enzyme without compromising methylation efficiency and requires less bench time. Herein, we describe the use of MAB-seq assay to evaluate the generation of 5-fC/5-caC in response to ATRA in mouse embryonic fibroblasts, using the hypermethylated in cancer 1 (Hic1) locus as a model system.
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Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Tretinoína
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Análise de Sequência de DNA
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5-Metilcitosina
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Fatores de Transcrição Kruppel-Like
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Fibroblastos
Tipo de estudo:
Prognostic_studies
Limite:
Animals
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article