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Efficient delivery of dsRNA into zona‐enclosed mouse oocytes and preimplantation embryos by electroporation
Author(s) -
Grabarek Joanna B.,
Plusa Berenika,
Glover David M.,
ZernickaGoetz Magdalena
Publication year - 2002
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/gene.10076
Subject(s) - electroporation , zona pellucida , blastocyst , embryo , green fluorescent protein , microbiology and biotechnology , biology , andrology , oocyte , zygote , transgene , embryogenesis , gene , genetics , medicine
Summary: 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. genesis 32:269–276, 2002. © 2002 Wiley‐Liss, Inc.