LncRNA H19介导表观遗传修饰重编程调控猪体细胞核移植胚胎发育机理的研究

体细胞核移植效率低限制了其应用。表观遗传修饰重编程不完全是克隆效率低的最主要原因。LncRNA参与表观遗传修饰，成为提高克隆效率研究的一个重要前沿。我们研究显示，lncRNA调控体细胞核移植重编程，而且，lncRNA H19参与克隆胚胎发育。但是，lncRNA H19调控克隆胚胎发育机理尚未明确。据此，本项目拟研究体细胞核移植过程中lncRNA H19表达特性；扰低或过表达后，检测克隆胚胎表观遗传修饰模式；单细胞转录组测序发掘候选基因，并调控其表达，明确lncRNA H19调控克隆胚胎发育的关键基因；分析lncRNA H19与表观遗传修饰酶互作及关键基因启动子区表观遗传修饰状态，构建lncRNA H19调控关键基因表达的表观遗传修饰网络，揭示lncRNA H19介导表观遗传修饰重编程调控克隆胚胎发育机理。该课题的探讨和阐明，将拓宽人们对体细胞核移植重编程机理的认识和推动克隆技术的广泛应用。

Low efficiency of somatic cell nuclear transfer limits its application. The main cause of low cloning efficiency is the incomplete epigenetic modification reprogramming. LncRNA is involved in the epigenetic modification, which becomes an important frontier to improve cloning efficiency. Our studies domenstrate that lncRNA regulates the reprogramming induced by somatic cell nuclear transfer, and lncRNA H19 is involved in the development of cloned embryos. However, the mechanism of lncRNA H19 regulating the development of cloned embryos is unclear. Accordingly, this topic will investigate the expression characteristics of lncRNA H19 during somatic cell nuclear transfer, study the epigenetic modification patterns of cloned embryos after lncRNA H19 knockdown or overexpression, apply single cell RNA sequencing to explore candidate genes, clarify the key genes associated with the cloned embryo development regulated by lncRNA H19 through regulating the expression of candidate genes, analyze the interaction between lncRNA H19 and epigenetic modification enzymes and the epigenetic modification status of key gene promoter, construct the epigenetic modification network of key gene expression regulated by lncRNA H19, and reveal the mechanism underlying the development of cloned embryos regulated by lncRNA H19 mediated epigenetic modification reprogramming. The discussion and clarification of this topic will broaden the understanding of the reprogramming mechanism induced by somatic cell nuclear transfer and promote the wide application of cloning technology.