重组蛋白诱导猪iPSCs及其表观遗传重编程变化规律研究

项目名称： 重组蛋白诱导猪iPSCs及其表观遗传重编程变化规律研究

项目编号： No.31272442

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 张运海

作者单位： 安徽农业大学

项目金额： 82万元

中文摘要： 猪作为重要经济动物和人类医学研究的动物模型，其iPSCs在疾病诊治、动物育种保种、生物学基础研究等方面具有重要价值。病毒介导获取的iPSCs存在安全隐患，iPSCs质量不理想等都限制了iPSCs的应用。本研究拟在前期工作基础上，建立并完善重组蛋白诱导iPSCs的技术方案，创建高质量、安全的iPSCs，并揭示iPSCs产生期间表观遗传重编程动态变化规律。重点从重组蛋白、源头细胞等环节优化iPSCs诱导方案；以病毒介导的iPSCs为对照，检测iPSCs的细胞表型、基因组特征及iPSCs产生期间的DNA甲基化、组蛋白修饰和microRNA变化规律，监测iPSCs克隆胚体内外发育，深入评估iPSCs质量和发育多能性，发掘iPSCs特异的microRNAs，丰富iPSCs质量评价标准。研究有助于加深我们对细胞重编程的认识、促进iPSC技术的不断完善和造福人类医学及畜牧业等。

中文关键词： 诱导多能干细胞；始发态样；体细胞重编程；非编码RNA；

英文摘要： Pig is generally regarded as an economically important domestic animal and an ideal animal model for human medical research, and it's induced pluripotent stem cells (iPSCs) hold great promise for disease diagnosis and drug screening, animal breeding and conservation, biology basic research. However, innative risks of virus-mediated iPSCs generation method, limited source cell type, sup-optimal culture conditions, and unideal iPSCs quality limit the wide application of iPSCs. Based on our previous work, non-virus-mediated and non-genome- integration strategy, this study aims to establish and improve the generation of iPSCs by using recombinant proteins, in order to efficiently generating high-quality, safe iPSCs lines, and to reveal epigenetic dynamic changes during the reprogramming. We will combine the recombinant protein, the source cell type, and small molecule intervention to optimise iPSCs induction program. iPSCs generated by virus-mediated methods serve as control, we then will examine the phenotype, biology properties and genome characteristics of iPSCs induced by the recombinant proteins. Dynamic changes of DNA methylation, histone modifications and microRNAs during reprogramming will also be explored. We will monitor the in vivo and in vitro developmental fate of the iPSCs cloned embryo to assess strin

英文关键词： induced pluripotent stem cells；naive like；somatic cell reprogramming；non-doding RNA；