CircRNA通过miR-148a/ miR-10a激活BMP7在湖羊毛囊发育中分子机理研究

湖羊是我国特有的白色羔皮用绵羊品种，以生产水波状羔皮著称于世。羔皮花纹形成与毛囊发育密切相关，但目前对其分子机制的了解甚少。本项目基于前期对BMP7在毛囊中的功能等研究基础上，拟通过核质穿梭等实验，阐明miR-148a/10a特异性调控BMP7的作用机制；通过检测羊毛乳头细胞的细胞增殖等实验，来明确候选circRNA、miR-148a/10a的功能；利用miRNA pull down等实验，验证circRNA—miR-148a/10a的调控关系；利用CRISPR/Cas9技术分别在羊毛乳头细胞和湖羊体内进行miR-148a 和miR-10a敲除，验证其功能以及circRNA—miR-148a/10a—BMP7之间的调控关系。研究结果将揭示circRNA通过miR-148a/10a作用于BMP7在湖羊羔皮毛囊发育中的功能、表达调控机制，为挖掘、开发和保护湖羊羔皮种质资源提供理论依据。

Hu sheep, a unique sheep breed in China, is well known for its water-wave lambskin pattern. The formation of lambskin pattern was closely associated with the development of hair follicle, but currently its molecular mechanism underlying the development of hair follicle is unknown. Based on our previous studies on the functions of BMP7 in hair follicle development, this project intends to elaborate the effect mechanism of action of miR-148a/ miR-10a in specifically activating BMP7 through various experimental methods including nucleosome shuttle. The functions of candidated circRNA and miR-148a / miR-10a will be confirmed by monitoring the cell proliferation of wool papilla cells,etc. The regulatory effects on the circRNA-miR-148a/ miR-10a-BMP7 will be verified by miRNA pull-down experiments, etc. And the miR-148a and miR-10a will be knockout by CRISPR / Cas9 technology in wool papilla cells and Hu sheep to verify their functions and the relationship among circRNA-miR-148a/miR-10a-BMP7. The results of this study will reveal the role of circRNAs and functions and expression mechanism of BMP7 in the development of Hu sheep lambskin hair follicles , and will provide the theoretical evidence for the excavation, development and protection of lambskin germplasm resources of Hu sheep.