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Nucleic Acids Research 2004 32(12):3601-3606; doi:10.1093/nar/gkh697
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Published online 9 July 2004

Nucleic Acids Research, Vol. 32 No. 12 © Oxford University Press 2004; all rights reserved

Lack of homologous sequence-specific DNA methylation in response to stable dsRNA expression in mouse oocytes

Petr Svoboda, Paula Stein1, Witold Filipowicz and Richard M. Schultz1,*

Friedrich Miescher Institute for Biomedical Research, PO Box 2543, 4002 Basel, Switzerland and 1 Department of Biology, University of Pennsylvania, 415 South University Avenue, Philadelphia, PA 19104-6018, USA

* To whom correspondence should be addressed. Tel: +1 215 898 7869; Fax: +1 215 898 8780; Email: rschultz{at}sas.upenn.edu

Received March 22, 2004; Revised May 30, 2004; Accepted June 22, 2004

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.


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