Using somatic-cell nuclear transfer to study aging
Using somatic-cell nuclear transfer to study aging
复制标题
利用体细胞核移植研究衰老
DOI：
10.1007/978-1-62703-556-9_9
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Wakayama T.
中科院分区：
文献类型：
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作者：
Kishigami S;Lee AR;Wakayama T.
在哺乳动物中，单倍体细胞、卵子和精子受精后的二倍体基因组是独特的和不可复制的。这意味着产生的独特的二倍体基因组注定与个人的不可避免的死亡。自1997年首次报道克隆绵羊多莉以来，许多哺乳动物物种都已利用体细胞核移植（SCNT）成功克隆。在哺乳动物中成功的SCNT不仅使我们能够在没有生殖细胞的情况下繁殖后代，也就是说，“通过”一个独特的二倍体基因组，而且还解决了发育，核重编程和表观遗传记忆等有价值的生物学问题。成功的克隆还可以支持表观遗传重编程，其中老化时钟被重置或逆转。最近使用iPS细胞技术的工作已经探索了实用性，并导致了来自早衰样核纤层蛋白病的iPSC过早衰老的重演。因此，重编程工具也有望有助于研究生物年龄。然而，在大多数情况下，动物克隆的效率仍然很低，克隆胚胎中的重编程机制在很大程度上仍然不清楚。在这里，基于最新的进展，我们描述了一种改进的，更有效的小鼠克隆协议，使用组蛋白脱乙酰酶抑制剂（HDACis）和latrunculin A，这增加了生产克隆小鼠或建立ES细胞的成功率五倍。这种改进的克隆方法将提供一个强有力的工具，以解决许多问题，包括生物老化更容易和更低的成本。
In mammals, a diploid genome following fertilization of haploid cells, an egg, and a spermatozoon is unique and irreproducible. This implies that the generated unique diploid genome is doomed with the individual’s inevitable demise. Since it was first reported in 1997 that Dolly the sheep had been cloned, many mammalian species have been cloned successfully using somatic-cell nuclear transfer (SCNT). The success of SCNT in mammals enables us not only to reproduce offspring without germ cells, that is, to “passage” a unique diploid genome, but also to address valuable biological questions on development, nuclear reprogramming, and epigenetic memory. Successful cloning can also support epigenetic reprogramming where the aging clock is reset or reversed. Recent work using iPS cell technology has explored the practicality and led to the recapitulation of premature aging with iPSCs from progeroid laminopathies. As a result, reprogramming tools are also expected to contribute to studying biological age. However, the efficiency of animal cloning is still low in most cases and the mechanism of reprogramming in cloned embryos is still largely unclear. Here, based on recent advances, we describe an improved, more efficient mouse cloning protocol using histone deacetylase inhibitors (HDACis) and latrunculin A, which increases the success rates of producing cloned mice or establishing ES cells fivefold. This improved method of cloning will provide a strong tool to address many issues including biological aging more easily and with lower cost.