建立Spanx基因条件敲除小鼠模型及其对精子生育能力影响的研究

项目名称： 建立Spanx基因条件敲除小鼠模型及其对精子生育能力影响的研究

项目编号： No.31472054

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 动物学

项目作者： 范立青

作者单位： 中南大学

项目金额： 85万元

中文摘要： 课题组前期研究发现,在生育力低下的精子和圆头精子中有SPANX蛋白家族多种蛋白表达下调;并证实SPANX蛋白与精子的生育能力密切相关,但具体作用机理未知。人类SPANX基因在减数分裂之后表达,推测该基因与精子核和顶体的形成相关，影响精子生育能力。此外,该基因在多种肿瘤和早衰卵巢中表达异常,提示其可能涉及肿瘤发生和卵巢发育。课题拟采用CRPSPR/Cas9技术，构建小鼠同源基因的条件敲除模型，结合临床上该基因与妊娠结局的相关性，探讨其与生育能力的关系，明确SPANX基因的功能；通过分析基因敲除后细胞和分子水平的改变，结合蛋白质相互作用研究，探索其调控的分子作用网络和机理。本课题拟建立SPANX对男性生育力评估体系，为男性不育的临床诊断和治疗提供指导，也为男性避孕药提供新的候选靶标,将有助于预防男性不育、改善男性生殖健康;动物模型的建立也为研究卵巢发育和肿瘤发生提供新的技术平台。

中文关键词： Spanx基因；生育力差异；基因敲除小鼠；CRISPR/Cas9；蛋白质组学

英文摘要： The mouse Spanxn gene is located on the X chromosome and specifically expressed in testes. The homologous Spanxn gene in human is a SPANX gene family, which contains two subfamilies SPANX-A/D and SPANX-N. Human SPANX gene family is post-meiotic expressed and is thought to be involved in the formation of acrosome and sperm nucleus. In our previous study on Globozoospermia patient's sperm with homozygous mutation of PICK1 gene, we found that compared with normal sperm, the expression of several SPANX proteins were significantly reduced in patient' s sperm. Moreover, we have found the expression of SPANX proteins was also significantly reduced in the donors'sperm with low fertility(no pregnancy in 12 AID treantment cycles). The Western Blotting result proved that the SPANX proteins is correlated to the fertility. However, the precise biological function of the SPANX gene family remains unclear. Besides, the SPANX gene was also found in many tumor cells and such as testescular germ cell tumor , melanoma, and probably involved in ovary development. In this project, we intend to use mice as a model since mice have a single form of SPANX, Spanxn，which will simplify the study of SPANX function. We will study the time and spacial expression pattern of Spanxn during mouse development. We will also establish a Spanxn knockout mice model and implement whole genome expression profiling and proteome analysis to uncover Spanxn function. Combining all the data above and with the use of bioinforatics, we intend to elucidate the molecular mechanisim and regulatory network of Spanxn. This project will establish Spanxn gene knockout model mice. It is meaningful to reveal the mechanism of Spanxn gene in spermatogenesis, especially in spermiogenesis stage, but also provide new model for ovary development and oncology research. Thus it will provide a theoretical basis and guidance for the clinical treatment of male infertility and may help us to find candidate targets for male contraceptive drugs and new contraceptive approach, and understand the mechanic basis of ovary development and the tumorigenesis

英文关键词： Spanx gene;fecundity diversity;gene konckout;CRISPR/Cas9;proteomics