title: Lack of homologous sequence-specific DNA methylation in response to stable dsRNA expression in mouse oocytes.
abstract: Double-stranded RNA (dsRNA) induces sequence-specific mRNA degradation in most eukaryotic organisms via a conserved pathway known as RNA interference (RNAi). Post-transcriptional gene silencing by RNAi is also connected with transcriptional silencing of cognate sequences. In plants, this transcriptional silencing is associated with sequence-specific DNA methylation. To address whether this mechanism operates in mammalian cells, we used bisulfite sequencing to analyze DNA in mouse oocytes constitutively expressing long dsRNA against the Mos gene. Our data show that long dsRNA induces efficient Mos mRNA knockdown but not CpG and non-CpG DNA methylation of the endogenous Mos sequence in oocytes and early embryos. These data demonstrate that dsRNA does not directly induce DNA methylation in the trans form of this sequence in these mammalian cells.
authors.affiliation: Friedrich Miescher Institute for Biomedical Research, PO Box 2543, 4002 Basel, Switzerland.
authors.forename: Petr
authors.initials: P
authors.lastname: Svoboda
authors.forename: Paula
authors.initials: P
authors.lastname: Stein
authors.forename: Witold
authors.initials: W
authors.lastname: Filipowicz
authors.forename: Richard M
authors.initials: RM
authors.lastname: Schultz
mesh_terms: D000818:Animals; D019175:DNA Methylation; D015870:Gene Expression; D016258:Genes, mos; D051379:Mice; D008822:Mice, Transgenic; D009865:Oocytes / Q000378:metabolism; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D013447:Sulfites / Q000737:chemistry
chemical_list: D012330:RNA, Double-Stranded; D013447:Sulfites