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如何提高小鼠克隆技术的成功率。

How to improve the success rate of mouse cloning technology.

作者信息

Thuan Nguyen Van, Kishigami Satoshi, Wakayama Teruhiko

机构信息

Department of Animal Bioscience and Biotechnology, Konkuk University, South Korea.

出版信息

J Reprod Dev. 2010 Feb;56(1):20-30. doi: 10.1262/jrd.09-221a.

Abstract

It has now been 13 years since the first cloned mammal Dolly the sheep was generated from somatic cells using nuclear transfer (SCNT). Since then, this technique has been considered an important tool not only for animal reproduction but also for regenerative medicine. However, the success rate is still very low and the mechanisms involved in genomic reprogramming are not yet clear. Moreover, the NT technique requires donated fresh oocyte, which raises ethical problems for production of human cloned embryo. For this reason, the use of induced pluripotent stem cells for genomic reprogramming and for regenerative medicine is currently a hot topic in this field. However, we believe that the NT approach remains the only valid way for the study of reproduction and basic biology. For example, only the NT approach can reveal dynamic and global modifications in the epigenome without using genetic modification, and it can generate offspring from a single cell or even a frozen dead body. Thanks to much hard work by many groups, cloning success rates are increasing slightly year by year, and NT cloning is now becoming a more applicable method. This review describes how to improve the efficiency of cloning, the establishment of clone-derived embryonic stem cells and further applications.

摘要

自第一只通过体细胞核移植(SCNT)技术从体细胞克隆出的哺乳动物多利羊诞生至今已有13年。从那时起,这项技术不仅被视为动物繁殖的重要工具,也被看作再生医学的重要手段。然而,其成功率仍然很低,基因组重编程所涉及的机制也尚不清楚。此外,核移植技术需要捐赠新鲜的卵母细胞,这引发了人类克隆胚胎生产中的伦理问题。因此,利用诱导多能干细胞进行基因组重编程和再生医学目前是该领域的一个热门话题。然而,我们认为核移植方法仍然是研究生殖和基础生物学的唯一有效途径。例如,只有核移植方法能够在不进行基因改造的情况下揭示表观基因组的动态和全局修饰,并且它能够从单个细胞甚至冷冻的尸体中产生后代。由于许多研究团队的不懈努力,克隆成功率逐年略有提高,现在核移植克隆正成为一种更具实用性的方法。这篇综述描述了如何提高克隆效率、建立克隆来源的胚胎干细胞以及进一步的应用。

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