Combinational Treatment of Trichostatin A and Vitamin C Improves the Efficiency of Cloning Mice by Somatic Cell Nuclear Transfer.

Combinational Treatment of Trichostatin A and Vitamin C Improves the Efficiency of Cloning Mice by Somatic Cell Nuclear Transfer.
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DOI:
10.3791/57036
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发表时间:
2018-04-26
期刊:
Journal of visualized experiments : JoVE
影响因子:
--
通讯作者:
Anzai M
Anzai M
中科院分区:
其他
文献类型:
--
作者:
Azuma R;Miyamoto K;Oikawa M;Yamada M;Anzai M

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体细胞核移植(SCNT)提供了一个从供体细胞直接产生克隆动物的独特机会,它需要使用熟练的技术。此外,自从克隆动物,特别是小鼠的成功生产以来,克隆的效率一直很低。为了提高克隆效率,人们进行了很多尝试,trichostatin A (TSA)作为一种组蛋白去乙酰化酶抑制剂被广泛应用于提高克隆效率。在这里,我们报告了一种显著改进的小鼠克隆方法。这种体细胞核移植方法使用日本包膜血凝病毒(HVJ-E),操作方便。此外,用TSA和维生素C (VC)两种小分子与去离子牛血清白蛋白(dBSA)一起处理,对胚胎发育非常有效。该方法既不需要额外的注射,也不需要基因操作,因此提供了一种简单,适合实际使用的方法。这种方法可能成为研究人员从培养细胞中培育转基因动物的技术上可行的方法。此外,它可能是通过克隆拯救濒危动物的一种有用的方法。
Somatic cell nuclear transfer (SCNT) provides a unique opportunity to directly produce a cloned animal from a donor cell, and it requires the use of skillful techniques. Additionally, the efficiencies of cloning have remained low since the successful production of cloned animals, especially mice. There have been many attempts to improve the cloning efficiency, and trichostatin A (TSA), a histone deacetylase inhibitor, has been widely used to enhance the efficiency of cloning. Here, we report a dramatically improved cloning method in mice. This somatic cell nuclear transfer method involves usage of Hemagglutinating virus of Japan Envelope (HVJ-E), which enables easy manipulation. Moreover, the treatment using two small molecules, TSA and vitamin C (VC), with deionized bovine serum albumin (dBSA), is highly effective for embryonic development. This approach requires neither additional injection nor genetic manipulation, and thus presents a simple, suitable method for practical use. This method could become a technically feasible approach for researchers to produce genetically modified animals from cultured cells. Furthermore, it might be a useful way for the rescue of endangered animals via cloning.
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