鉴定影响产奶性状的主效基因及其调控机制对实施分子育种以加快遗传进展尤为重要。申请人所在团队前期利用RNA-Seq和small RNA-Seq技术检测了奶牛不同泌乳阶段肝脏组织差异表达的mRNA、lncRNA和miRNA，构建了以miRNA为中心的ceRNA调控网络，并通过功能富集分析发现了12个与乳脂性状相关的关键候选基因。本研究旨在进一步利用双荧光素酶报告系统、过表达、RNA干扰和RNA结合蛋白免疫沉淀等功能基因组学技术在细胞水平上确定ceRNA网络中miRNA、lncRNA和mRNA之间的相互作用，并检测其对乳脂合成相关基因的作用，以揭示ceRNA在奶牛肝脏组织中对乳脂性状形成的调控机制。

Identification of major genes affecting milk production traits and their regulatory mechanisms is particularly important for accelerating genetic progress in dairy cattle. In the previous study, we used RNA-Seq and small RNA-Seq to obtain liver transcriptome of dairy cattles in different lactation stages, and identified the differentially expressed mRNA, lncRNA, and miRNA across the three lactation periods. We have built ceRNA regulatory networks via miRNA to reveal the regulatory mechanisms for milk production traits in liver of dairy cattle, and the critical genes of ceRNA network in milk fat synthesis have be enriched by biological functional analysis. In this study, we will validate the interactions among RNAs in ceRNA regulatory network at cell level to confirm its regulation using dual-luciferase reporter system, overexpression or interference of RNAs, and RNA binding protein immunoprecipitation. Furthermore, we will detect the influence of ceRNA on the expression of critical genes in milk fat synthesis to reveal the regulatory mechanism of ceRNA in dairy cattle liver.