To improve our understanding of the molecular mechanisms underlying early embryo development, further characterization of gene activity in oocytes and embryos is urgently required. The transition from the two-cell to four-cell stage is particularly important in pre-implantation embryonic development, as it involves transcriptional reprogramming and cellular differentiation. In this study, we used a 7.4 K cDNA microarray to screen mRNA transcript levels in the pre-implantation mouse embryo. Real-time PCR was used to confirm microarray data. We profiled 7,410 genes and identified 4,562 genes that were differentially expressed in the pre-implantation embryo. We selected a total of 248 genes with significant expression changes that are functionally involved in the two-cell and two-cell block embryo. Of these genes, 114 were down-regulated and the remainder (n=134) were up-regulated in the two-cell embryo. This study provides a developmental map of a large number of genes in the pre-implantation mouse embryo with particular emphasis on gene expression in the two-cell embryo and two-cell block embryo. Further investigations based on this data will provide a better understanding of the effects of various external conditions and may facilitate comparative analysis of pre-implantation development in other mammalian species, including human.