Identification of genes aberrantly expressed in mouse embryonic stem cell-cloned blastocysts
Identification of genes aberrantly expressed in mouse embryonic stem cell-cloned blastocysts
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DOI：
10.1095/biolreprod.107.064634
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发表时间：
2008-04-01
影响因子：
3.6
通讯作者：
Kono, Tomohiro
中科院分区：
文献类型：
--
作者：
Jincho, Yuko;Sotomaru, Yusuke;Kono, Tomohiro
在发育过程中，克隆胚胎经常在胚胎发生的任何阶段经历胚胎停滞，导致各种形态异常。胚胎克隆程序造成的长期影响将在出生后表现为早死、肥胖、各种功能障碍等等。尽管进行了广泛的研究，但影响克隆胚胎发育特征的参数仍不清楚。本研究进行了广泛的基因表达分析，以筛选胚胎干细胞克隆囊胚中异常表达的基因簇。cDNA减法文库的差异筛选显示，克隆囊胚中有224个差异表达基因，其中85个通过BLAST搜索确定为具有广泛功能的已知基因。为了证实它们的差异表达，采用实时荧光定量PCR对单个囊胚进行了基因表达分析。在大多数胚胎干细胞克隆胚胎中，Skp1a、Canx、Ctsd、Timd2和Psrnc6基因显著上调，Aqp3、Ak3l1、Rhot1、Sf3b3、Nid1、mt-Rnr2、mt-Nd1、mt-Cytb和mt-Co2基因显著下调。我们的研究结果表明，由胚泡期各种基因的异常表达引起的多种功能障碍的异常高频率与克隆胚胎的发育停滞和各种其他疾病有关。
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 Psrnc6 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.