Recent Advance in Genome Editing-Based Gene Modification in Pigs
Recent Advance in Genome Editing-Based Gene Modification in Pigs
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DOI：
10.5772/intechopen.88022
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发表时间：
2019-07
期刊：
影响因子：
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通讯作者：
Masahiro Sato;K. Miyoshi;H. Kawaguchi;E. Inada;I. Saitoh;A. Tanimoto
中科院分区：
文献类型：
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作者：
Masahiro Sato;K. Miyoshi;H. Kawaguchi;E. Inada;I. Saitoh;A. Tanimoto
最近，包括ZFN、TALENs和CRISPR/Cas9系统在内的一系列基因组编辑技术已经实现了包括猪在内的各种生物体的内源性靶基因的基因修饰，这对于农业和生物医学研究非常重要。由于其简单的基因敲除应用和易用性，CRISPR/Cas9现在在全球范围内普遍使用。这个过程中最重要的方面是选择用于将基因组编辑组件传递到胚胎的方法。在早期阶段，经常采用将这些组分[单向导RNA（sgRNA）+Cas9的DNA/mRNA]显微注射到受精卵的细胞质和/或细胞核中。然而，这种方法总是与嵌合胚胎的产生有关，其中基因组编辑和未编辑的细胞混合在一起。为了避免这种嵌合体问题，在与Cas9蛋白混合的sgRNA（称为核糖核蛋白（RNP））存在下对受精卵进行体外电穿孔现在被频繁使用。这篇综述提供了基因组编辑猪生产的历史背景，并介绍了目前关于如何在体细胞核移植衍生的胚胎中诱导基因组编辑的研究，这些胚胎已用正常细胞核重建。
Recently, a series of genome editing technologies including ZFNs, TALENs, and CRISPR/Cas9 systems have enabled gene modification in the endogenous target genes of various organisms including pigs, which are important for agricultural and biomedical research. Owing to its simple application for gene knockout and ease of use, the CRISPR/Cas9 is now in common use worldwide. The most important aspect of this process is the selection of the method used to deliver genome editing components to embryos. In earlier stages, zygote microinjection of these components [single guide RNA (sgRNA) + DNA/mRNA for Cas9] into the cytoplasm and/ or nuclei of a zygote has been frequently employed. However, this method is always associated with the generation of mosaic embryos in which genome-edited and unedited cells are mixed together. To avoid this mosaic issue, in vitro electroporation of zygotes in the presence of sgRNA mixed with Cas9 protein, referred to as a ribonucleoprotein (RNP), is now in frequent use. This review provides a historical background of the production of genome-edited pigs and also presents current research concerning how genome editing is induced in somatic cell nuclear transfer-derived embryos that have been reconstituted with normal nuclei.