猪长链非编码RNA-ED1与miR-21的互作及其对骨骼肌生长发育的调控

The interaction between Long non-coding RNA (lncRNA) and microRNA is a hot spot in current research, and sponge effect is a major way of the interaction forms. We have identified lncRNA-ED1(long non-coding RNA- Embryonic Differentiation 1) was a muscle-specific long non-coding RNA, miR-21 was involved in the regulation of myogenesis in pigs. Therefore, we inferred that lncRNA-ED1 may regulate skeletal muscle development by functioning as miR-21’s sponge. In this study, we will employ lean-type landrace, fat-type Tongcheng pig and analyze the negative feedback of lncRNA-ED1 and miR-21 in totally 28 developmental time points of the two different types. We will explore the correlation between lncRNA-ED1 and proliferation、differentiation、 apoptosis of the primary muscle cells. Meanwhile, promoter confusion experiment, RIP and Pull down are further used to reveal the functions of lncRNA-ED1 as miR-21’s sponge. Our study will provide theoretical basis for genetics of skeletal muscle development in different types of pigs and new gene material in pig breeding.

长链非编码RNA（lncRNA）与microRNA互作是当前研究热点，海绵体效应是互作的一种主要方式。本组前期发现一个新lncRNA-ED1(long non-coding RNA- Embryonic Differentiation 1)与猪骨骼肌发育相关，同时揭示了miR-21参与猪骨骼肌发育调控，在此基础上深入探讨lncRNA-ED1对miR-21海绵体效应及其对猪骨骼肌生长发育的调控。本项目拟综合分析lncRNA-ED1和miR-21在瘦肉型长白猪、脂肪型通城猪肌肉发育过程28个时间节点的负反馈互作。以猪原代肌细胞为模型系统研究lncRNA-ED1对骨骼肌细胞增殖、分化、凋亡影响。进一步通过启动子融合构建、RIP及Pull down 技术鉴定lncRNA-ED1对miR-21海绵体效应。揭示骨骼肌发育及中外猪种骨骼肌表型差异分子遗传学基础，为我国猪产肉性状遗传改良提供新的基因素材。

长链非编码RNA（lncRNA）与microRNA互作是当前研究热点，申请者前期利用深度测序的方法鉴定了猪胚胎期90天（E90）和出生后100天（D100）骨骼肌中表达的microRNAs，发现miR-21在胚胎期90天（E90）的表达量是出生后100天（D100）的17倍，利用小鼠成肌细胞系C2C12过表达或抑制miR-21，首次发现并证实了miR-21能显著促进成肌细胞的增殖；随后利用生物信息学方法预测并鉴定出miR-21的一个靶标基因TGFBI，揭示了miR-21通过调控TGFB/AKT-mToR信号通路调控骨骼肌的发育。后期通过高通量芯片数据分析发现一个新的lncRNA-ED1(long non-coding RNA- Embryonic Differentiation 1)在猪骨骼肌发育中与miR-21的表达呈现负相关。本项目中，申请者分析了高通量芯片表达结果及差异，鉴定了lncRNA-ED1和miR-21在瘦肉型长白猪、脂肪型通城猪肌肉发育过程28个时间节点的负反馈互作。以小鼠成肌细胞和猪原代肌细胞为模型系统研究lncRNA-ED1对骨骼肌细胞增殖和分化的影响，揭示了lncRNA-ED1在中外猪种骨骼肌表型差异分子遗传学基础。