SUV39H1/2基因表达抑制对转基因克隆猪胚胎早期发育潜能及外源基因表达水平的影响研究

转基因克隆动物在农业和医学领域有着广泛的应用前景，目前制约该技术应用和发展的主要问题是生产效率不高，这主要与体细胞核移植效率低、外源基因整合率及基因表达沉默等因素有关。项目选择在体细胞重编程和调控基因转录活性两方面均具有重要作用的SUV39H1/2基因为研究对象，以稳定表达外源基因的猪成纤维细胞系为材料，采用RNA干扰技术及核移植方法研究SUV39H1/2基因表达调控对转基因克隆猪胚胎早期发育潜力及外源基因表达水平的影响。通过分析重构胚合子基因组激活及早期胚胎发育相关、DNA甲基化和组蛋白乙酰化等重要基因的表达水平，以及外源基因启动子甲基化变化初步阐明其作用机理。课题研究可为进一步解析SUV39H1/2基因在体细胞重编程及基因转录调控功能、完善转基因克隆动物生产技术体系打下基础。

Transgenic cloning animal has a broad application in agriculture and medicine fields, but the low production efficiency of current transgenic animal technology limits its application and development. The main reason is related to prospects, such as the low efficiency of somatic cell nuclear transfer technology, the low integration rate of exogenous gene and gene expression silence. SUV39H1/2 genes, which play an important role both in the somatic cell reprogramming and the regulation of gene transcription activity, are selected to be studied. The porcine fibroblast lines with stable expression of exogenous gene are selected as materials, RNA interference technology and nuclear transfer method are used to explore the effect of down regulating the expression of SUV39H1/2 genes on the developmental potential of porcine transgenic cloned embryos and the transgene expression levels. The expression of genes related to the zygote genomic activation, the early embryonic development, DNA methylation and histone acetylation are analyzed, the changes of promoter methylation level of exogenous gene in transgenic cloned pig embryos were detected, which would clarify the mechanism of SUV39H1/2 genes. The results would help us to further know the function of SUV39H1/2 genes in somatic cell reprogramming and regulating gene translation activity, and lay the foundation to improve the production efficiency of transgenic cloning animal.