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CRISPR/Cas9介导的基因组编辑中的嵌合现象。

Mosaicism in CRISPR/Cas9-mediated genome editing.

作者信息

Mehravar Maryam, Shirazi Abolfazl, Nazari Mahboobeh, Banan Mehdi

机构信息

Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.

Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran; Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran.

出版信息

Dev Biol. 2019 Jan 15;445(2):156-162. doi: 10.1016/j.ydbio.2018.10.008. Epub 2018 Oct 22.

Abstract

The CRISPR/Cas9 system is a rapid, simple, and often extremely efficient gene editing method. This method has been used in a variety of organisms and cell types over the past several years. However, using this technology for generating gene-edited animals involves a number of obstacles. One such obstacle is mosaicism, which is common in founder animals. This is especially the case when the CRISPR/Cas9 system is used in embryos. Here we review the pros and cons of mosaic mutations of gene-edited animals caused by using the CRISPR/Cas9 system in embryos. Furthermore, we will discuss the mechanisms underlying mosaic mutations resulting from the CRISPR/Cas9 system, as well as the possible strategies for reducing mosaicism. By developing ways to overcome mosaic mutations when using CRISPR/Cas9, genotyping for germline gene disruptions should become more reliable. This achievement will pave the way for using the CRISPR technology in the research and clinical applications where mosaicism is an issue.

摘要

CRISPR/Cas9系统是一种快速、简单且通常极为高效的基因编辑方法。在过去几年中,该方法已被应用于多种生物体和细胞类型。然而,将这项技术用于生成基因编辑动物存在诸多障碍。其中一个障碍是嵌合体现象,这在奠基动物中很常见。当在胚胎中使用CRISPR/Cas9系统时尤其如此。在此,我们综述了在胚胎中使用CRISPR/Cas9系统导致基因编辑动物发生嵌合突变的利弊。此外,我们将讨论CRISPR/Cas9系统导致嵌合突变的潜在机制,以及减少嵌合体现象的可能策略。通过开发在使用CRISPR/Cas9时克服嵌合突变的方法,种系基因破坏的基因分型应会变得更加可靠。这一成果将为在嵌合体现象成为问题的研究和临床应用中使用CRISPR技术铺平道路。

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