Manipulating transgenes using a chromosome vector
Manipulating transgenes using a chromosome vector
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DOI：
10.1093/nar/gkp058
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发表时间：
2009-04-01
影响因子：
14.9
通讯作者：
Okazaki, Tsuneko
中科院分区：
文献类型：
--
作者：
Ikeno, Masashi;Suzuki, Nobutaka;Okazaki, Tsuneko
最近的技术进步使我们能够可视化局部染色质结构的组织和动态，然而，染色质组织调节基因调控的综合机制知之甚少。我们设计了一种人类人工染色体载体，该载体允许使用将染色质结构递送到从人类到鱼类的不同细胞系中的方法来操纵转基因。这种方法能够分析从头构建，表观遗传维持和特定基因的染色质结构的变化。分别从小鼠胚胎干细胞和CHO细胞中的裸DNA建立了人STAT3的表达和抑制结构。在抑制性结构中向胚胎干细胞递送STAT3导致STAT3活化，伴随着DNA甲基化的变化。这种操纵具有特定染色质结构的单个基因的技术可用于应用生物学，包括干细胞科学和再生医学。
Recent technological advances have enabled us to visualize the organization and dynamics of local chromatin structures; however, the comprehensive mechanisms by which chromatin organization modulates gene regulation are poorly understood. We designed a human artificial chromosome vector that allowed manipulation of transgenes using a method for delivering chromatin architectures into different cell lines from human to fish. This methodology enabled analysis of de novo construction, epigenetic maintenance and changes in the chromatin architecture of specific genes. Expressive and repressive architectures of human STAT3 were established from naked DNA in mouse embryonic stem cells and CHO cells, respectively. Delivery of STAT3 within repressive architecture to embryonic stem cells resulted in STAT3 activation, accompanied by changes in DNA methylation. This technology for manipulating a single gene with a specific chromatin architecture could be utilized in applied biology, including stem cell science and regeneration medicine.