title: RNAi-based methods for gene silencing in mouse oocytes.
abstract: RNA interference (RNAi) is an evolutionary conserved gene-silencing pathway that can be efficiently utilized as a tool to study gene function. RNAi is initiated by long double-stranded RNAs (dsRNAs), which are processed into small duplexes called small-interfering RNAs (siRNAs). In turn, these duplexes target mRNAs for degradation in a sequence-specific manner. Mouse oocytes, unlike most mammalian cell types, lack an interferon response to long dsRNA. Moreover, they are a rare example of a mammalian cell type with a robust endogenous RNAi pathway. For these reasons microinjection of either long dsRNAs or siRNAs results in efficient, sequence-specific gene silencing. Here, we describe a protocol for preparation and microinjection of long dsRNA into mouse oocytes.
authors.affiliation: Department of Biology, University of Pennsylvania, Philadelphia, PA, USA, steinpau@sas.upenn.edu.
authors.forename: Paula
authors.initials: P
authors.lastname: Stein
authors.forename: Petr
authors.initials: P
authors.lastname: Svoboda
authors.forename: Richard M
authors.initials: RM
authors.lastname: Schultz
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D005260:Female; D055029:Inverted Repeat Sequences; D051379:Mice; D008845:Microinjections / Q000379:methods*; D009865:Oocytes / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics* / Q000302:isolation & purification; D048348:Reverse Transcription
chemical_list: D012330:RNA, Double-Stranded