增强子RNA（eRNAs）是由活性增强子转录产生的一种新型的非编码RNA，它对靶基因的表达具有重要的调控作用。由于猪骨骼肌增强子的信息较少，eRNAs鉴定的技术难度较大，缺乏有效的细胞模型等原因，目前eRNAs调控猪骨骼肌发育的分子机制仍不清楚。为了解决这一问题，本项目以猪胚胎期骨骼肌纤维为研究对象，首先利用CUT&Tag技术进行增强子的定位，通过GRO-seq方法检测新生RNA，并联合多组学数据对eRNAs进行鉴定和分类；然后，以猪骨骼肌卫星细胞为模型，通过RNAi方法验证MyoD对eRNAs转录的促进作用，同时利用CRISPR/Cas9系统验证eRNAs对MyoD基因表达的调控作用，并获取eRNAs结合蛋白及其关键结合位点信息来探究eRNAs和转录因子MyoD的正反馈作用关系。本项目既有助于推进猪骨骼肌发育调控的分子机制研究，更可以为猪产肉性状的遗传改良提供有效的分子标记。

Enhancer RNA (eRNAs) is a new type of noncoding RNA transcribed by active enhancer, which plays an important role in regulating the transcription activity of target genes. However, at present, the molecular mechanism of the regulation of porcine skeletal muscle development by eRNAs is still unclear. The main reasons are as follows: the information of pig skeletal muscle enhancer is deficient, the technology of identification of eRNAs is difficult, and there is no effective cell model. In order to solve this problem, this project takes the skeletal muscle fiber of pig embryo as the research object. Cut & Tag technology is used to locate the enhancer, and then GRO-seq method is applied in detecting nascent RNA. Multiple omics data are integrated to identify and classify the eRNAs. On this basis, taking the porcine skeletal muscle satellite cell as the model, it explores the positive feedback relationship between the eRNAs and the transcription factor MyoD. RNAi method is used to verify the role of MyoD in promoting the transcription of eRNAs. Then, CRISPR / Cas9 is used to verify its role in the transcription of MyoD gene. Finally, the binding protein of eRNAs is detected and its key binding sites are analyzed. It is not only helpful to promote the research of molecular mechanism of pig skeletal muscle development regulation, but also provide effective molecular markers for genetic improvement of pig meat production traits.