绵羊克隆囊胚差异表达microRNA 的筛选及其功能研究

体细胞核移植（克隆）技术已在农业、物种保护、医疗等领域显示出诱人的应用前景。然而，克隆胚胎发育异常及效率低一直是制约克隆技术发展应用的瓶颈。microRNA（miRNA）是新发现的一类非编码RNA，具有调控动物胚胎发育、细胞增殖和分化的功能。然而miRNA与核移植克隆胚胎发育异常的关系，迄今仍然不清楚。前期研究中，本课题组发现与绵羊体内受精囊胚相比，核移植克隆囊胚中部分miRNA表达量显著上调。因此推测异常表达的miRNA可能导致克隆胚胎发育异常，这可能是一种导致克隆胚胎发育异常的新机制。为证实这一假说，本研究拟采用体细胞核移植，绵羊超排冲胚，Illumina高通量测序等技术研究绵羊克隆囊胚与体内受精囊胚miRNA表达谱差异；并分析这些差异表达miRNA的靶基因及其在胚胎发育中的功能，从miRNA角度阐明绵羊体细胞克隆胚胎发育异常的分子机制，为提高体细胞克隆效率提供新手段和依据。

Somatic cell nucleus transfer (SCNT) technology show a good application prospect in agriculture, species conservation and medicine. However, developmental abnormalities of cloned embryo always restricted development and application of SCNT. MicroRNA (miRNA)，a new non-coding RNA，play an important role in cell proliferation and differentiation during mammalian development. But the associations between miRNA and developmental abnormalities of cloned embryo are lacking. In our earlier study, we found that some miRNAs were upregulated in cloned blastula compared to in-vivo fertilized blastula. Thus, we speculate that the aberrated expression of miRNA result in developmental abnormalities of cloned embryo, which may be a mechanism of aberrated development of cloned embryo. To confirm our speculation, we want to study microRNA expression profiling of cloned blastula and in-vivo fertilized blastula by SCNT, superovulation, embryo flushing and Illumina technology. Differential expression miRNA will be determined by bioinformatics. Theirs target gene and function will be analyzed in development of cloned embryo.The study will determined the role of miRNA in developmental abnormalities of cloned embryo,which will provide a new method for increase cloned animal rate.