HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 75/6/891    most recent biolreprod.106.054213v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Amanai, M. Articles by Perry, A. C.F. Search for Related Content PubMed PubMed Citation Articles by Amanai, M. Articles by Perry, A. C.F. Agricola Articles by Amanai, M. Articles by Perry, A. C.F.

Injection of Mammalian Metaphase II Oocytes with Short Interfering RNAs to Dissect Meiotic and Early Mitotic Events¹

Laboratory of Mammalian Molecular Embryology, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan

ABSTRACT

The manipulation of mammalian metaphase II (mII) oocytes has illuminated the mechanisms of fertilization and early embryogenesis and is central to nuclear transfer. Although RNA interference (RNAi) would greatly facilitate this type of manipulation, its application to mature, developmentally competent mII oocytes has not been evaluated. We report efficient RNAi by the injection of short interfering RNAs (siRNAs) into mII oocytes. The levels of the target mRNA and corresponding protein were rapidly and efficiently reduced. The siRNAs were effective when injected in the subnanomolar to nanomolar range and induced concurrently RNAi of multiple targets, revealing the kinetic parameters of RNAi in mII oocytes. Coinjection of sperm with siRNA functionally abolished the transcripts in the resultant blastocysts and in cloned embryos into which siRNA was coinjected during somatic cell nuclear transfer. The RNAi method was used to dissect the early mitotic roles of meiotic regulators, which suggests that CDC20 is essential for the first mitotic division, while EMI1 and EMI2 are not essential for this process. Our results show that siRNA injection of oocytes confers temporal control of RNAi in the analysis and manipulation of key processes in mammalian meiosis and early embryogenesis.

early development, embryo, fertilization, gamete biology

¹Supported by grant-in-aid (no. C90-53010) from the Ministry of Education, Sports, Science, and Technology of Japan.

Correspondence: ² Tony Perry, Laboratory of Mammalian Molecular Embryology, RIKEN Center for Developmental Biology, 2-2-3 Minatojima Minamimachi, Chuo-ku, Kobe 650-0047 Japan. FAX: 81 78 306 3144; e-mail: tony{at}cdb.riken.jp

This article has been cited by other articles:

				N. Yoshida, M. Amanai, T. Fukui, E. Kajikawa, M. Brahmajosyula, A. Iwahori, Y. Nakano, S. Shoji, J. Diebold, H. Hessel, et al. Broad, ectopic expression of the sperm protein PLCZ1 induces parthenogenesis and ovarian tumours in mice Development, November 1, 2007; 134(21): 3941 - 3952. [Abstract] [Full Text] [PDF]