小鼠卵子重构介导的生殖性半克隆研究

当前辅助生殖技术迅猛发展，但却无法使配子缺乏的夫妇获得自己遗传学上的后代。通过生殖性半克隆技术，即体细胞核移植技术获得克隆配子，与配偶正常配子结合生育自己遗传特征的后代是绝大多数患者夫妇的愿望，也是生殖医学的重大突破。项目申请人既往研究中首创性的建立了 “人造精子”介导的半克隆技术。然而，通过克隆卵子获得半克隆囊胚及动物研究尚未见报道。项目申请人基于既往研究基础，在预实验中实现领域突破，通过体细胞核移植技术获得克隆卵子，并在体外与正常精子受精，成功获得小鼠半克隆囊胚。本项目拟在预实验的基础上通过重编程因子促进体细胞重编程过程来建立更加高效的卵子重构生殖性半克隆技术并获得半克隆动物，同时构建胚胎干细胞系从基因组整倍性及表观遗传探究其安全性及有效性；从重构卵的纺锤体形成、同源染色体配对分离以及参与的关键蛋白和基因探索其发生机制。本项目的研究将会加深对配子发育的理解，为临床不孕治疗提供新的思路。

Currently, assisted reproductive technology (ART) develops rapidly. However, there is no way to sovle this problem for couples who lack for gametes. The clonal gametes obtained through somatic cell nuclear transfer （SCNT）can fertiled with normal gametes to give birth to offspring, namely reproductive semi-cloned technology, which can enable couples without gametes to give birth to offspring with their own genetic characteristics. It is desirable by majority of infertile couples and will be also a huge breakthrough in reproductive medicine. In previous research, the project applicant established "artificial sperm" mediated semi-cloning technology for the first time. The establishment, research and application of semi-cloned technology mediated by "artificial sperm" have been very mature and in-depth. However, the study of semi-cloned blastocysts or animal obtained by somatic cell egg reconstructed in vitro has not been reported yet. On the basis of previous research, we made a huge breakthrough in the field in the preliminary experiment by obtaining cloned eggs through somatic cell nuclear transfer technology, which was fertilized with normal sperm in vitro and successfully obtained mouse semi-cloned blastocysts. On the basis of the pre-experiment, this project intends to establish a more efficient reproductive semi-cloned technology for egg reconstruction and obtain semi-cloned animals by promoting the process of somatic cell reprogramming through reprogramming factors, and meanwhile, to construct embryonic stem cell lines to explore its safety and effectiveness from genomic euploidy and epigenetics. Moreover, we will further figure out the mechanism through spindle formation, homologous chromosome division of reconstructed egg genome and key gene in this process. This study will deepen the understanding of gamete development, provide a new idea for clinical infertility treatment, and have a profound impact on reproductive medicine.