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This Article Full Text Full Text (PDF) All Versions of this Article: 78/4/568    most recent biolreprod.107.064634v2 biolreprod.107.064634v1 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 Google Scholar Articles by Jincho, Y. Articles by Kono, T. PubMed PubMed Citation Articles by Jincho, Y. Articles by Kono, T. Agricola Articles by Jincho, Y. Articles by Kono, T.

Identification of Genes Aberrantly Expressed in Mouse Embryonic Stem Cell-Cloned Blastocysts¹

Department of Bioscience,³ Tokyo University of Agriculture, Tokyo 156-8502, Japan Natural Science Center for Basic Research and Development,⁴ Hiroshima University, Hiroshima 734-8551, Japan

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.

cDNA subtraction, early development, embryo, ES cell clone, gene expression, gene regulation, nuclear transfer

¹Supported by grants-in-aid from the Ministry of Education, Science, Sports and Culture of Japan and from the Ministry of Agriculture, Forestry and Fisheries of Japan (Development of stable production technology of cloned animals by somatic cell nuclear transfer) to T.K.

Correspondence: ²Tomohiro Kono, Department of Bioscience, Tokyo University of Agriculture 1-1-1, Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan. FAX: 81 3 5477 2543; e-mail: tomohiro{at}nodai.ac.jp