normal dna methylation dynamics in dicer1-deficient mouse embryonic stem cells正常的dna甲基化动力学dicer1-deficient老鼠胚胎干细胞.pdfVIP

Normal DNA Methylation Dynamics in DICER1-Deficient Mouse Embryonic Stem Cells 1,2 1 1 1 1 1 Jonathan Ip , Paul Canham , K. H. Andy Choo , Yoshimi Inaba , Shelley A. Jacobs , Paul Kalitsis , 1 3 3 3 4 1 Deidre M. Mattiske , Jane Ng , Richard Saffery , Nicholas C. Wong , Lee H. Wong , Jeffrey R. Mann * 1Theme of Genetic Disorders, Murdoch Childrens Research Institute, The Royal Children’s Hospital, Parkville, Victoria, Australia, 2 Department of Zoology, The University of Melbourne, Melbourne, Victoria, Australia, 3 Theme of Cell Biology, Development, and Disease, Murdoch Childrens Research Institute, The Royal Children’s Hospital, Parkville, Victoria, Australia, 4 Department of Biochemistry and Molecular Biology, Monash University, Melbourne, Victoria, Australia Abstract Reduced DNA methylation has been reported in DICER1-deficient mouse ES cells. Reductions seen at pericentric satellite repeats have suggested that siRNAs are required for the proper assembly of heterochromatin. More recent studies have postulated that the reduced methylation is an indirect effect: the loss of Mir290 cluster miRNAs leads to upregulation of the transcriptional repressor RBL2 that targets the downregulation of DNA methyltransferase (Dnmt) genes. However, the observations have been inconsistent. We surmised that the inconsistency could be related to cell line ‘‘age,’’ given that DNA methylation is lost progressively with passage in DNMT-deficient ES cells. We therefore subjected Dicer12/ 2 ES cells to two experimental regimes to rigorously test the level of functional DNMT activity. First, we cultured th