title: Efficient delivery of dsRNA into zona-enclosed mouse oocytes and preimplantation embryos by electroporation.
abstract: Conditions for the electroporation of mouse oocytes and preimplantation embryos have been optimised by following the incorporation of rhodamine labeled dextran. This procedure includes a step to weaken but not remove the zona pellucida that helps achieve good survival. This approach has been applied to introduce double-stranded RNA for c-mos into oocytes and green fluorescent protein (GFP) into transgenic GFP-expressing embryos at the 1- and 4-cell stages. In both cases we were able to observe sequence-specific interference with the expression of the target gene--a failure of oocytes to arrest at metaphase II and a loss in the green fluorescence of embryos by the morula or blastocyst stages. These effects could be observed in multiple oocytes or embryos allowed to develop together following electroporation.
authors.affiliation: Cyclacel Ltd., Polgen Division, Babraham Bioincubator, Babraham Cambridge, United Kingdom.
authors.forename: Joanna B
authors.initials: JB
authors.lastname: Grabarek
authors.forename: Berenika
authors.initials: B
authors.lastname: Plusa
authors.forename: David M
authors.initials: DM
authors.lastname: Glover
authors.forename: Magdalena
authors.initials: M
authors.lastname: Zernicka-Goetz
mesh_terms: D000818:Animals; D001483:Base Sequence; D001755:Blastocyst / Q000166:cytology / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D017931:DNA Primers; D018274:Electroporation / Q000379:methods; D016258:Genes, mos; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D009865:Oocytes / Q000502:physiology*; D017231:Proto-Oncogene Proteins c-mos / Q000235:genetics*; D012330:RNA, Double-Stranded / Q000235:genetics*; D011994:Recombinant Proteins / Q000032:analysis; D015203:Reproducibility of Results; D015044:Zona Pellucida / Q000502:physiology*
chemical_list: D017931:DNA Primers; D008164:Luminescent Proteins; D012330:RNA, Double-Stranded; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D017231:Proto-Oncogene Proteins c-mos