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Cell ; 163(1): 218-29, 2015 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-26406378

RESUMO

Mammalian DNA methylation plays an essential role in development. To date, only snapshots of different mouse and human cell types have been generated, providing a static view on DNA methylation. To enable monitoring of methylation status as it changes over time, we establish a reporter of genomic methylation (RGM) that relies on a minimal imprinted gene promoter driving a fluorescent protein. We show that insertion of RGM proximal to promoter-associated CpG islands reports the gain or loss of DNA methylation. We further utilized RGM to report endogenous methylation dynamics of non-coding regulatory elements, such as the pluripotency-specific super enhancers of Sox2 and miR290. Loci-specific DNA methylation changes and their correlation with transcription were visualized during cell-state transition following differentiation of mouse embryonic stem cells and during reprogramming of somatic cells to pluripotency. RGM will allow the investigation of dynamic methylation changes during development and disease at single-cell resolution.


Assuntos
Metilação de DNA , Análise de Célula Única , Animais , Ilhas de CpG , Metilases de Modificação do DNA/metabolismo , Células-Tronco Embrionárias , Elementos Facilitadores Genéticos , Humanos , Camundongos , MicroRNAs/metabolismo , Regiões Promotoras Genéticas , Fatores de Transcrição SOXB1/metabolismo
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