LncEPC5通过编码小肽调节原始生殖细胞形成的机制解析

原始生殖细胞（PGCs）已成为禽类种质资源保护和遗传改造的工具细胞，然而因其体内分离数量较少、体外诱导效率低而导致PGCs来源不足，极大限制其应用，根本原因在于不清楚PGCs发生的机制。因此解析PGCs发生机制十分紧迫和必要。课题组前期研究发现LncEPC5能够通过调节BMP4和编码小肽EPC5的方式调节鸡PGCs形成，且EPC5能够与DDX5发生互作。据此我们提出假说：LncEPC5通过编码的小肽EPC5调节BMP4或DDX5并影响下游关键分子是鸡PGCs形成的主要原因。为验证上述假说，本项目拟在明确EPC5的表达规律和功能基础上，通过ChIP-seq和双荧光素酶报告系统分析EPC5是否通过调节BMP4参与鸡PGCs的形成；通过RIP测序和RNA-Pull Down解析EPC5与DDX5的互作机制以及DDX5调节鸡PGCs形成的分子机制。研究结果为提高PGCs诱导效率提供技术支撑。

Primordial germ cells (PGCs) can be used as a tool cell for the protection of poultry germplasm resources and genetic modification. However, the insufficient sources of PGCs due to its small number isolated in vivo and low induction efficiency in vitro have greatly limited its application. The fundamental reason is that the mechanism of PGCs formation is not clear. Therefore, it is urgent and necessary to analyze the mechanism of PGCs. In the previous research, we found that LncEPC5 could regulate chicken PGCs formation via changing BMP4 expression and coding small peptide EPC5 by using chicken as animal model. Further, it turned out that EPC5 was interacted with DDX5. According to the results above, we came up with a hypothesis that the main reason of chicken PGCs development might cause by the small peptide EPC5 which was coded by LncEPC5 regulate BMP4 or DDX5 expression and affect the expression of downstream key molecule. In order to elucidate the hypothesis above, this program plan to clarify the regularity of EPC5 expression and study the function of EPC5 during chicken PGCs development via different research strategies both in vitro and in vivo; Analysis by Dual luciferase reporter assay to find out whether EPC5 participate in the regulation process of PGCs development via changing BMP promoter to affect BMP expression; Through RIP sequence and RNA-Pull down to interpret the molecular mechanism of PGCs formation. The research results would provide the technique support for improving PGCs induction efficiency.