摘要
不完全的表观遗传重编程是造成转基因克隆动物效率低下的主要原因,组蛋白修饰作为表观遗传修饰的一个重要部分,可以直接影响克隆胚胎的发育和外源基因的表达情况。TSA(Trichostatin A)作为一种组蛋白去乙酰化抑制剂,可以改变组蛋白的乙酰化水平,促进表观遗传重编程,提高克隆动物的效率。同时TSA能改变染色质结构,使转录因子易于与DNA序列结合,促进外源基因的表达。文章确定了TSA处理转基因猪成纤维细胞和核移植胚胎的最佳条件,分别为250 nmol/L、24 h和40 nmol/L、24 h,通过进一步正交实验发现,TSA同时处理供体细胞和克隆胚胎可以显著的促进核移植胚胎的体外发育。此外,无论TSA处理转基因猪成纤维细胞或核移植胚胎,都可以提高外源基因的表达水平。
Uncompleted epigenetic reprogramming is attributed to the low efficiency of producing transgenic cloned animals.Histone modification associated with epigenetics can directly influence the embryo development and transgene expression.Trichostatin A（TSA）,as an inhibitor of histone deacetylase,can change the status of histone acetylation,im-prove somatic cell reprogramming,and enhance cloning efficiency.TSA prevents the chromatin structure from being con-densed,so that transcription factor could binds to DNA sequence easily and enhance transgene expression.Our study estab-lished the optimal TSA treatment on porcine donor cells and cloned embryos,250 nmol/L,24 h and 40 nmol/L,24 h,re-spectively.Furthermore,we found that both the cloned embryo and the donor cell treated by TSA resulted in the highest development efficiency.Meanwhile,TSA can improve transgene expression in donor cell and cloned embryo.In summary,TSA can significantly improve porcine reconstructed embryo development and transgene expression.
出处
《遗传》
CAS
CSCD
北大核心
2011年第7期749-756,共8页
Hereditas（Beijing)
基金
转基因生物新品种培育科技重大专项(编号:2008ZX08006-002
2009ZX08006-001B)
东北农业大学"创新团队"发展计划项目资助
关键词
TSA
体细胞核移植
胚胎发育
外源基因表达
猪
TSA
somatic cell nuclear transfer
embryo development
transgene expression
pig
