title: Identification of genes aberrantly expressed in mouse embryonic stem cell-cloned blastocysts.
abstract: During development, cloned embryos often undergo embryonic arrest at any stage of embryogenesis, leading to diverse morphological abnormalities. The long-term effects resulting from embryo cloning procedures would manifest after birth as early death, obesity, various functional disorders, and so forth. Despite extensive studies, the parameters affecting the developmental features of cloned embryos remain unclear. The present study carried out extensive gene expression analysis to screen a cluster of genes aberrantly expressed in embryonic stem cell-cloned blastocysts. Differential screening of cDNA subtraction libraries revealed 224 differentially expressed genes in the cloned blastocysts: eighty-five were identified by the BLAST search as known genes performing a wide range of functions. To confirm their differential expression, quantitative gene expression analyses were performed by real-time PCR using single blastocysts. The genes Skp1a, Canx, Ctsd, Timd2, and Psmc6 were significantly up-regulated, whereas Aqp3, Ak3l1, Rhot1, Sf3b3, Nid1, mt-Rnr2, mt-Nd1, mt-Cytb, and mt-Co2 were significantly down-regulated in the majority of embryonic stem cell-cloned embryos. Our results suggest that an extraordinarily high frequency of multiple functional disorders caused by the aberrant expression of various genes in the blastocyst stage is involved in developmental arrest and various other disorders in cloned embryos.
authors.affiliation: Department of Bioscience, Tokyo University of Agriculture, Tokyo 156-8502, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Jincho
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Sotomaru
authors.forename: Manabu
authors.initials: M
authors.lastname: Kawahara
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Ono
authors.forename: Hidehiko
authors.initials: H
authors.lastname: Ogawa
authors.forename: Yayoi
authors.initials: Y
authors.lastname: Obata
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Kono
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000737:chemistry*; D003001:Cloning, Molecular; D019976:Cloning, Organism; D018076:DNA, Complementary / Q000032:analysis / Q000737:chemistry; D053595:Embryonic Stem Cells / Q000737:chemistry*; D005260:Female; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D016133:Polymerase Chain Reaction
chemical_list: D018076:DNA, Complementary