肉用山羊circ_0009659通过miR-23b-3p/KLF8轴调控肌内脂肪细胞分化的分子机制研究

肌内脂肪细胞分化对提高山羊肉产量及品质具有重要意义。我们前期发现肉用山羊circ_0009659在肌内脂肪细胞分化前后表达差异极显著，干扰后抑制肌内脂肪细胞脂滴积聚，且主要定位于细胞质，结合实验室前期研究结果推测其可能通过miR-23b-3p/KLF8轴调控肉用山羊肌内脂肪细胞分化。本项目拟通过功能获得/缺失实验明确肉用山羊circ_0009659对肌内脂肪细胞分化的调控作用；利用RNA pull-down、RNA-FISH及荧光素酶报告系统等技术鉴定circ_0009659/miR-23b-3p/KLF8之间的竞争关系并揭示相互调控作用；进一步利用RNA-seq及恢复实验确定circ_0009659通过miR-23b-3p/KLF8轴调控肌内脂肪细胞分化的下游关键信号通路，系统阐明circ_0009659的分子调控机制，研究结果为提高山羊肉产量及改善肉品质提供新的理论基础和科学依据。

Intramuscular adipocyte differentiation is of great significance in improving meat yield and quality in meat goat. In our previous study, we found that the circ_0009659 expression of meat goat was significant difference between preadipocytes and mature adipocyte, and the interference of circ_0009659 inhibited the accumulation of lipid droplets in intramuscular adipocytes. The circ_0009659 is mainly localized in the cytoplasm. Combined with our previous results, it is speculated that circ_0009659 may regulate the differentiation of goat intramuscular adipocytes through the miR-23b-3p/KLF8 axis. This project intends to clarify the regulatory effects of meat goat circ_0009659 on the differentiation of intramuscular adipocytes through function gain/loss experiments, as well as, to identify the competitive relationships among circ_0009659/miR-23b-3p/KLF8, and reveal their mutual regulatory effects using techniques such as RNA pull-down, RNA-FISH and luciferase reporting system. Further, using RNA-seq and recovery experiments, we will determine the key down-stream signal pathways of circ_0009659 in regulating intramuscular adipocyte differentiation via the miR-23b-3p/KLF8 axis, in order to elucidate the molecular regulatory mechanism of circ_0009659. The results from this research will provide a new theoretical basis and scientific evidences for improving goat meat yield and quality.