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Publication : Transgenic RNAi in mouse oocytes: a simple and fast approach to study gene function.

First Author  Stein P Year  2003
Journal  Dev Biol Volume  256
Issue  1 Pages  187-93
PubMed ID  12654301 Mgi Jnum  J:82704
Mgi Id  MGI:2654413 Doi  10.1016/s0012-1606(02)00122-7
Citation  Stein P, et al. (2003) Transgenic RNAi in mouse oocytes: a simple and fast approach to study gene function. Dev Biol 256(1):187-93
abstractText  Double-strand RNA (dsRNA)-mediated posttranscriptional gene silencing, also known as RNA interference (RNAi), is a powerful tool to inhibit gene expression in several experimental model systems, including Arabidopsis, Caenorhabditis, and Drosophila. We previously described that the microinjection of Mos dsRNA into fully grown mouse oocytes results in the specific degradation of Mos mRNA in a time- and concentration-dependent manner. We report here a transgenic RNAi approach that is suitable to study gene function during mouse oocyte development and differentiation. The oocyte-specific Zp3 promoter was used to drive the expression of a long hairpin dsRNA ( approximately 500 bp) targeting Mos mRNA. Transgenic founder animals appeared healthy, but while males were fertile, females were not, in accordance with the known Mos null phenotype. The amount of Mos mRNA in the transgenic F(1) females was reduced by >90%, whereas there was no decrease in the nontargeted tissue plasminogen activator (Plat) mRNA. Moreover, the maturation-associated increase in mitogen-activated protein (MAP) kinase activity was not observed, and the metaphase II eggs underwent spontaneous parthenogenetic activation, thus recapitulating the Mos null phenotype. This approach provides a powerful method to study the functions of any oocyte-synthesized gene during oocyte development and early embryogenesis.
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