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CRISPR/Cas9(clustered regulatory interspaced short palindromic repeats/CRISPR-associated protein 9)系统在活细胞中能精确有效地进行基因组编辑,被广泛应用在生命科学的各个领域中。相比原有的基因编辑技术,CRISPR/Cas9基因编辑技术程序简易、操作方便灵活,极大简化了遗传工程的操作,提高了基因编辑效率。文章回顾了CRISPR/Cas9基因编辑技术的发展历程,阐述了其作用机制及在哺乳动物猪上的应用。通过sgRNA的设计原理、作用机制对CRISPR/Cas9技术存在的问题进行分析,试图提高其打靶效率,降低脱靶效应,为优化CRISPR/Cas9的设计及应用提供参考。
The CRISPR / Cas9 (clustered regulatory interspaced short palindromic repeats / CRISPR-associated protein 9) system is capable of genome-accurate editing in living cells and is widely used in various fields of life sciences. Compared with the original gene editing technology, CRISPR / Cas9 gene editing technology is simple and easy to operate, which greatly simplifies the operation of genetic engineering and improves the efficiency of gene editing. The paper reviews the development of CRISPR / Cas9 gene editing technology and expounds its mechanism of action and its application in mammalian pigs. Through the design principle and mechanism of sgRNA, the problems existing in CRISPR / Cas9 technology are analyzed in order to improve its targeting efficiency and reduce the off-target effect, so as to provide reference for optimizing the design and application of CRISPR / Cas9.