International Mammalian Genome Society

The 16th International Mouse Genome Conference (2002)


W Kimber
Developmental Genomics & Aging Section, Laboratory of Genetics; National Institute on Aging, NIH

Puri N, Borgmeyer C, Ritter D, Seidman M, Ko M

Mouse preimplantation development encompasses embryo development from fertilization up until implantation.  Many complex developmental events take place during this period, including the transition from maternal to zygotic transcription, establishment of embryonic axes, and formation of the blastocyst, the first mammalian differentiation event.  Using a multi-faceted approach, we aim to identify and study genes with key roles in these processes.  Our study utilizes the unique resource of the NIA cDNA clone set, which contains many novel, full length ESTs representing all mouse preimplantation stages.  This clone set provides the basis for microarray experiments enabling us to study global gene expression, and assemble gene expression profiles.  In addition, we are using EST frequency in individual libraries to identify genes with restricted expression profiles, implying a role at specific stages of preimplantation development.  These studies are complemented with functional assays including gene knockouts, in vitro expression analysis, and gene ‘knock-down’ experiments using antisense technology.  As a prelude to the application of antisense oligonucleotides technology to gene expression studies in preimplantation embryos in vitro, we have evaluated the effect of a number of oligonucleotide sugar modifications on target specificity and nuclease resistance.  We show that 2‘-methoxyethoxy modified oligonucleotides were significantly more effective in suppressing gene expression than either conventional phosphorothioate oligonucleotides or oligonucleotides bearing a morpholino modification.  These studies lay the foundation for a systematic evaluation of the role of large numbers of genes in preimplantation development.

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