Dynamic visualization of DNA methylation in cell cycle genes during iPSC cardiac differentiation.

Autor:	Li N; Division of Cardiology, Department of Medicine, University of Washington, Seattle, WA, USA.; State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China., Nguyen BT; Department of Anatomy, Biochemistry and Physiology, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, HI, USA.; Center for Cardiovascular Research, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, HI, USA., Stitt EA Jr; Department of Anatomy, Biochemistry and Physiology, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, HI, USA.; Developmental and Reproductive Biology Graduate Program, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, HI, USA., Zhang Z; Division of Cardiology, Department of Medicine, University of Washington, Seattle, WA, USA., MacLellan WR; Division of Cardiology, Department of Medicine, University of Washington, Seattle, WA, USA., Zhang Y; Division of Cardiology, Department of Medicine, University of Washington, Seattle, WA, USA.; Department of Anatomy, Biochemistry and Physiology, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, HI, USA.; Center for Cardiovascular Research, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, HI, USA.; Developmental and Reproductive Biology Graduate Program, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, HI, USA.
Jazyk:	angličtina
Zdroj:	Epigenomics [Epigenomics] 2024 Dec-Dec; Vol. 16 (23-24), pp. 1407-1414. Date of Electronic Publication: 2024 Nov 27.
DOI:	10.1080/17501911.2024.2430171
Abstrakt:	Background: Existing analyses with conventional assays have generated significant insights into static states of DNA methylation but were unable to visualize the dynamics of epigenetic regulation. Materials & Results: We utilized a genomic DNA methylation reporter (GMR) system carrying Snrpn minimal promoter and CpG regions of Cdk1 (Cyclin-dependent kinase 1) or Sox2 (SRY-Box Transcription Factor 2). Mouse Sox2 GMR iPSCs rapidly lost fluorescent reporter signal upon the induction of cardiac differentiation. Cdk1 GMR reporter signal was strong in undifferentiated iPSCs, and gradually decreased during cardiomyocyte differentiation. RT-qPCR and pyrosequencing demonstrated that the reduction of Sox2 and Cdk1 was regulated by hypermethylation of their promoters' CpG regions during cardiac differentiation. Conclusion: The GMR reporter system can be useful for monitoring real-time epigenetic DNA modification at single-cell resolution.
Databáze:	MEDLINE
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