Selective reduction of dormant maternal mRNAs in mouse oocytes by RNA interference.
Specific mRNA degradation mediated by double-stranded RNA (dsRNA), which is termed RNA interference (RNAi), is a useful tool with which to study gene function in several systems. We report here that in mouse oocytes, RNAi provides a suitable and robust approach to study the function of dormant maternal mRNAs. Mos (originally known as c-mos) and tissue plasminogen activator (tPA, Plat) mRNAs are dormant maternal mRNAs that are recruited during oocyte maturation; translation of Mos mRNA results in the activation of MAP kinase. dsRNA directed towards Mos or Plat mRNAs in mouse oocytes effectively results in the specific reduction of the targeted mRNA in both a time- and concentration-dependent manner. Moreover, dsRNA is more potent than either sense or antisense RNAs. Targeting the Mos mRNA results in inhibiting the appearance of MAP kinase activity and can result in parthenogenetic activation. Mos dsRNA, therefore, faithfully phenocopies the Mos null mutant. Targeting the Plat mRNA with Plat dsRNA results in inhibiting production of tPA activity. Finally, effective reduction of the Mos and Plat mRNA is observed with stoichiometric amounts of Mos and Plat dsRNA, respectively.
Svoboda
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P
Department of Biology, University of Pennsylvania, Philadelphia, PA 19104-6018, USA.
Stein
P
P
Hayashi
H
H
Schultz
R M
RM
D000818:Animals; D016203:CDC2 Protein Kinase / Q000378:metabolism; D019926:Cyclin B / Q000378:metabolism; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism*; D009866:Oogenesis; D014176:Protein Biosynthesis / Q000187:drug effects*; D017231:Proto-Oncogene Proteins c-mos / Q000235:genetics*; D012330:RNA, Double-Stranded / Q000494:pharmacology*; D023121:RNA, Messenger, Stored / Q000378:metabolism*; D010959:Tissue Plasminogen Activator / Q000096:biosynthesis / Q000235:genetics*
D019926:Cyclin B; D012330:RNA, Double-Stranded; D023121:RNA, Messenger, Stored; D017231:Proto-Oncogene Proteins c-mos; D016203:CDC2 Protein Kinase; D020928:Mitogen-Activated Protein Kinases; D010959:Tissue Plasminogen Activator