Identification of inappropriately reprogrammed genes by large-scale transcriptome analysis of individual cloned mouse blastocysts.
Although cloned embryos generated by somatic/embryonic stem cell nuclear transfer (SECNT) certainly give rise to viable individuals, they can often undergo embryonic arrest at any stage of embryogenesis, leading to diverse morphological abnormalities. In an effort to gain further insights into reprogramming and the properties of SECNT embryos, we performed a large-scale gene expression profiling of 87 single blastocysts using GeneChip microarrays. Sertoli cells, cumulus cells, and embryonic stem cells were used as donor cells. The gene expression profiles of 87 blastocysts were subjected to microarray analysis. Using principal component analysis and hierarchical clustering, the gene expression profiles were clearly classified into 3 clusters corresponding to the type of donor cell. The results revealed that each type of SECNT embryo had a unique gene expression profile that was strictly dependent upon the type of donor cells, although there was considerable variation among the individual profiles within each group. This suggests that the reprogramming process is distinct for embryos cloned from different types of donor cells. Furthermore, on the basis of the results of comparison analysis, we identified 35 genes that were inappropriately reprogrammed in most of the SECNT embryos; our findings demonstrated that some of these genes, such as Asz1, Xlr3a and App, were appropriately reprogrammed only in the embryos with a transcriptional profile that was the closest to that of the controls. Our findings provide a framework to further understand the reprogramming in SECNT embryos.
Fukuda
Atsushi
A
Department of Bioscience, Tokyo University of Agriculture, Tokyo, Japan.
Cao
Feng
F
Morita
Shinnosuke
S
Yamada
Kaori
K
Jincho
Yuko
Y
Tane
Shouji
S
Sotomaru
Yusuke
Y
Kono
Tomohiro
T
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism*; D019976:Cloning, Organism; D015536:Down-Regulation; D020869:Gene Expression Profiling; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D015854:Up-Regulation