牛胚胎发育相关精子特异miRNAs的鉴定和功能研究

项目名称： 牛胚胎发育相关精子特异miRNAs的鉴定和功能研究

项目编号： No.31472094

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 畜牧学与草地科学

项目作者： 王勇胜

作者单位： 西北农林科技大学

项目金额： 87万元

中文摘要： 越来越多的证据表明精子受精过程中带入胚胎的miRNAs可能对胚胎的发育和表观重编程修饰起着重要的调控作用。本项目拟利用新一代高通量测序技术构建牛成熟精子、卵母细胞及皮肤成纤维细胞miRNAs文库，筛选精子特异表达miRNAs。通过生物信息学分析和双荧光素酶报告试验揭示关键精子特异miRNAs的预测靶基因及其生物调控作用；利用功能缺失策略研究精子特异关键miRNAs对受精胚胎发育、基因表达及表观重编程的影响；利用功能获得试验研究精子特异关键miRNAs对体细胞克隆胚胎的质量及克隆效率的影响。系统揭示精子特异miRNAs的生物学功能，阐明其对胚胎发育和（或）表观重编程的调控机制，扩展对早期胚胎发育及表观重编程调控的认识，为改进体细胞克隆技术和体外诱导干细胞技术提供新的思路。

中文关键词： 胚胎发育；重编程；精子miRNAs；体细胞核移植；分子机制

英文摘要： Accumulated evidences have shown that sperm miRNAs might play important roles in embryonic development and nuclear epigenetic reprogramming. The aims of the present objective are to construct miRNA libraries of bovine matured sperm, oocyte and skin fibroblast using the next generation of high-throughput sequencing technique, and screen the developmentally important miRNAs specially expressed in matured sperm. The target genes of key miRNAs are predicted by bioinformatics analysis and dual luciferase reporter system. The effects of key miRNAs on the development, gene expression, and epigenetic reprogramming of early embryo are studied by function-deprivation strategy. The effects of key miRNAs on the efficiency of the somatic cell nuclear transfer are evaluated by function-obtaining strategy. We hope that the project would reveal the biological function of the sperm-specific miRNAs systematically, clarify their regulatory mechanisms on development and (or) epigenetic reprogramming of early embryo, expand the knowledge of regulation of early embryonic development and epigenetic reprogramming, and provide new ideas for improving somatic cell cloning and in vitro induced stem cell technology.

英文关键词： embryonic development;reprogramming;sperm miRNAs;somatic cell nuclear transfer;Molecular mechanisms