鸡脂类合成主要发生在肝脏中。Vanin-1是一种泛酰巯基乙胺酶，在哺乳动物中对肝脏脂类代谢具有重要的调控作用，而它在鸡脂类代谢中的功能尚不清楚。本项目拟采用RACE并结合生物信息学和荧光素酶报告基因的方法以及ChIP和EMSA、定点突变等技术，确定鸡Vanin-1基因的启动子结构、转录因子精确结合位点，弄清该基因的转录调控机制；通过生物信息学筛选出能与该基因配对结合的miRNA，并用双荧光素酶报告基因进行验证以研究其转录后调控机制；采用过表达和CRISPRI基因干扰技术，分别在鸡LMH细胞系和原代鸡肝细胞中过表达和敲低该基因，高通量测序检测肝细胞中差异表达基因，并检测细胞及培养基中与脂质代谢相关的生化指标，运用qRT-PCR和Western Blot检测脂质代谢相关基因的表达量，综合分析Vanin-1在鸡肝脏脂类代谢中的功能。研究结果将有助于认识鸡肝脏脂类代谢和控制脂肪沉积的分子机理。

De novo synthesis of fatty acids in chickens occurs in liver. Vanin-1 is a pantetheinase that plays important roles in lipid metabolism in mammals, but its roles in lipid metabolism in chickens are still unknown. In this study, the RACE method, the bioinformatics, reporter gene assay, chromatin immunoprecipitation (ChIP) , electrophoretic mobility shift assay (EMSA), as well as site-directed mutagenesis will be combined to confirm the structure of Vanin-1 promoter, binding sites of transcription factors, and to understand regulatory mechanism of Vanin-1 gene in chickens. Through bioinformatics, miRNAs were screened which can directly bind to Vanin-1 gene 3’UTR. Then, the mechanism of post-transcription regulation of Vanin-1 gene was verified by co-transfection and dual-luciferase reporter assays. Using clustered regularly interspaced short palindromic repeat sequences interference (CRISPRi) system to knockdown the expression level of Vanin-1 in primary chicken hepatocytes, and differentially expressed genes in the chicken hepatocytes were detected by high throughput sequencing technology. Furthermore, detection of the expression of genes related to lipid metabolism was performed in chicken hepatocytes after over-expressing Vanin-1 by fluorescence quantitative PCR and Western Blot. Meanwhile the biochemical parameters related to lipid metabolism will be detected in the chicken hepatocytes with knockdown or over expression of Vanin-1 and in culture medium by biochemical methods. Taken together, the roles of Vanin-1 gene in chicken lipid metabolism will be concluded comprehensively. The results of this study will help to understand the molecular mechanism of lipid metabolism and control of fat deposition in chicken liver.