CRISPR/Cas9系统及其在鸡基因组编辑中应用的研究进展

doi:10.16431/j.cnki.1671-7236.2018.06.021

摘要/Abstract

摘要：

近年来，基因组编辑技术的快速发展促进了畜禽重要经济性状功能基因的研究及分子标记辅助育种的应用。由成簇间隔短回文重复序列（clustered regularly interspaced short palindromic repeats，CRISPR）与CRISPR相关蛋白9（CRISPR-associated protein 9，Cas9）组成的CRISPR/Cas9技术，是近年来新开发的一种高效的基因组编辑技术。与锌指核酸酶技术（zinc finger endonuclease，ZFN）和转录激活因子样效用物核酸酶技术（transcription activator-like effector nuclease，TALEN）相比，CRISPR/Cas9技术具有操作简单、成本低、效率高和适用范围广等优点，已广泛应用于人类细胞、果蝇、斑马鱼、小鼠、猪、鸡等动物及植物的基因组定点修饰，在畜禽基因组编辑和应用中具有广阔的应用前景。CRISPR/Cas9系统具有抵御外源病毒或质粒入侵的获得性免疫功能，其由sgRNA通过碱基互补配对识别靶DNA序列，引导Cas9特异性切割，实现基因组特定DNA片段的敲除、敲入或替换。鸡作为第一个完成全基因组测序的农业动物，不仅是一种重要的经济动物，也是生物学和分子遗传学基础研究中的重要模式生物。作者将从CRISPR/Cas9的研究历程、分类、作用机制、鸡基因组编辑中的应用及脱靶效应等方面进行综述，为利用该技术进行鸡基因功能的研究、重要经济性状的遗传改良和分子定向育种提供重要的参考。

关键词: CRISPR/Cas9系统; 基因编辑; 鸡; 脱靶效应; 分子育种

Abstract:

In recent years,the rapid development of genome editing promotes studies of functional genes associated with economically important traits and applications of molecular marker assisted breeding in livestock and poultry.Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) is a newly developed and highly efficient tool for genome editing.Compared with zinc finger endonuclease (ZFN) and transcription activator-like effector nuclease (TALEN),it is characteristic of simple operation,low cost,highly efficient and wide application,which has been widely used in human cells,Drosophila melanogaster,zebrafish,mice,pig,chicken and plants for gene targeting.The CRISPR/Cas9 is the cutting edge of genome editing and broad application in livestock and poultry breeding.The CRISPR/Cas9 system is an acquired immune system which fights against invasion of exogenous virus or plasmids.It has been modified as efficient genome editing technique that can be directed to knockout,knock-in,or substitution of target genes by single guide RNA (sgRNA) to identify foreign DNA and target it for inactivation by nuclease-mediated cleavage.The chicken represents the first agricultural animal to complete its genome sequenced,which serves as an economically important animal and an ideal animal model in biological and molecular genetics studies.In this review,we summarize the evolutionary history,classification,functional mechanism,its application in chicken genome editing,and off-target effect,which will provide important references for gene function studies,genetic improvement of important economic traits,and molecular directed breeding in chickens.

Key words: CRISPR/Cas9 system; genome editing; chicken; off-target effect; molecular breeding

中图分类号:

邱桂如, 王政, 何婷婷, 耿照玉, 金四华. CRISPR/Cas9系统及其在鸡基因组编辑中应用的研究进展[J]. 《中国畜牧兽医》, 2018, 45(6): 1590-1598.

QIU Guiru, WANG Zheng, HE Tingting, GENG Zhaoyu, JIN Sihua. Research Advances on CRISPR/Cas9 System and Its Application in Genome Editing in Chickens[J]. , 2018, 45(6): 1590-1598.

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