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Stem Cell Reports. 2014 Jun 3;2(6):910-24. doi: 10.1016/j.stemcr.2014.05.008. eCollection 2014 Jun 3.

Heterochromatin dynamics during the differentiation process revealed by the DNA methylation reporter mouse, MethylRO.

Author information

1
Center for Genetic Analysis of Biological Responses, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita 565-0871, Japan.
2
Department of Advanced Medical Initiatives, JST-CREST, Faculty of Medicine, Kyushu University, Fukuoka 812-8582, Japan.
3
Graduate School of Medicine, Osaka University, Suita 565-0871, Japan.
4
Research and Development Center, Fuso Pharmaceutical Industries, Ltd., Osaka 536-8523, Japan.
5
Graduate School of Frontier Biosciences, Osaka University, Suita 565-0871, Japan.

Erratum in

  • Stem Cell Reports. 2014 Jul 8;3(1):216.

Abstract

In mammals, DNA is methylated at CpG sites, which play pivotal roles in gene silencing and chromatin organization. Furthermore, DNA methylation undergoes dynamic changes during development, differentiation, and in pathological processes. The conventional methods represent snapshots; therefore, the dynamics of this marker within living organisms remains unclear. To track this dynamics, we made a knockin mouse that expresses a red fluorescent protein (RFP)-fused methyl-CpG-binding domain (MBD) protein from the ROSA26 locus ubiquitously; we named it MethylRO (methylation probe in ROSA26 locus). Using this mouse, we performed RFP-mediated methylated DNA immunoprecipitation sequencing (MeDIP-seq), whole-body section analysis, and live-cell imaging. We discovered that mobility and pattern of heterochromatin as well as DNA methylation signal intensity inside the nuclei can be markers for cellular differentiation status. Thus, the MethylRO mouse represents a powerful bioresource and technique for DNA methylation dynamics studies in developmental biology, stem cell biology, as well as in disease states.

PMID:
24936475
PMCID:
PMC4050349
DOI:
10.1016/j.stemcr.2014.05.008
[Indexed for MEDLINE]
Free PMC Article

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