HHEX基于miR-204-5p/CBL/Notch信号通路特异性调控猪体长性状的分子机理研究

猪体长是重要的经济性状之一，与脊椎数和单节椎体长直接相关。目前猪单节椎体长的研究鲜有报道。申请人前期研究发现显著差异的椎体长是影响五指山猪与大白猪体长的重要因素之一。已有的组学数据表明miR-204-5p-HHEX-CBL互做表达与猪体长性状密切相关。本项目拟进一步探明HHEX介导的miR-204-5p/CBL/Notch信号通路调控猪体长性状的分子机理。项目将在细胞和组织水平分析HHEX在软骨发育过程中的表达模式，通过RNA-seq和Pathway-Array分析精准确定HHEX所介导的Notch通路作用途径，利用ChIP-seq、CoIP和Pulldown等技术分析HHEX与miR-204-5p和CBL互作激活Notch信号通路的作用模式，最后利用基因编辑小鼠验证HHEX通过Notch信号通路调控软骨发育及体长性状的分子机制。上述机制的解析对于猪体长性状的遗传改良具有重要意义。

Body length of pigs is one of the important economic traits, which is directly related to the number of vertebrae and the length of single centrum. Few studies about the single centrum length of pigs have been currently reported. Our previous research has found the significant difference in the length of single centrum is one of the important reasons for explaining the body length difference between Wuzhishan pig and Large White pig. And our previous omics results demonstrated that the interaction expression of miR-204-5p-HHEX-CBL is closely related to the body length of pigs. Based on previous multi-omics data, the molecular mechanism of HHEX regulating body length of pigs via miR-204-5p-CBL-Notch signaling pathway will be a potential research topic for elucidating the above body length differences between pig breeds. The study will analyze the expression pattern of HHEX during chondrogenic development. Then fine determination of the role of HHEX mediated Notch signaling pathway by using RNA-seq and Pathway-Array. Furthermore, investigating the mechanism that HHEX activates Notch signaling pathway through interaction with miR-204-5p and CBL by using ChIP-seq、CoIP and Pulldown techniques. Finally, the specific regulation mechanism of pig body length by HHEX via the miR-204-5p-CBL-Notch Signaling Pathway will be verified by transgenic mouse. This study is very important for the genetic improvement of pig body length.