Efficient delivery of dsRNA into zona-enclosed mouse oocytes and preimplantation embryos by electroporation |
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Authors: | Grabarek Joanna B Plusa Berenika Glover David M Zernicka-Goetz Magdalena |
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Affiliation: | Cyclacel Ltd., Polgen Division, Babraham Bioincubator, Babraham Cambridge, United Kingdom. |
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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. |
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Keywords: | mouse oocytes preimplantation embryos RNA interference electroporation |
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