CRISPR/Cas9系统介导基因组编辑的研究进展

doi:10.11843/j.issn.0366-6964.2014.10.001

摘要/Abstract

摘要：

由规律成簇间隔短回文重复序列（Clustered regularly interspaced short palindromic repeats，CRISPR）以及CRISPR相关蛋白9（CRISPR-associated protein 9，Cas9），即CRISPR/Cas9系统改造的第3代人工核酸内切酶，与锌指核酸内切酶(Zinc finger endonuclease，ZFN)以及类转录激活因子效应物核酸酶(Transcription activator-like effector nuclease，TALEN)相比，具有构建方法简单快捷、突变效率高、成本低廉、适用范围广等许多优点，已成为一种热门的基因组编辑工具。该技术已成功应用于细菌、人类细胞、斑马鱼、小鼠、猪、食蟹猴以及多种植物的基因组精确修饰，是最具有临床与应用前景的基因治疗技术之一。但是CRISPR/Cas9系统目前也面临着脱靶率高、特异性差的难题，这无疑是CRISPR/Cas9系统应用于基础研究和临床治疗的最大的挑战。本文从CRISPR/Cas9系统的发现、分类、作用机制以及CRISPR/Cas9基因编辑系统在基因定点修饰方面的研究进展进行介绍，希望能够为畜牧领域的科研工作者提供参考。

关键词: 人工核酸内切酶, 基因编辑, 基因打靶, CRISPR/Cas9, RNA指导的核酸内切酶

Abstract:

Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) constituted the CRISPR/Cas9 genome editing system，which provides a more efficient way for gene targeting than zinc finger endonuclease (ZFN) and transcription activator-like effector nuclease (TALEN).It is becoming a hot spot in the field of genome editing，and successfully applied to gene targeting of bacteria，human cells，zebra fish，mouse，pig，cynomolgus monkey and several plants.However，the high off-target rate and poor specificity of CRISPR/Cas9 technology hindered its application in medical treatment.Here，the research progress in the structure，classification，function，and mechanism of the CRISPR/Cas9 system is reviewed.

Key words: engineered endonuclease, genome editing, gene targeting, CRISPR/Cas9, RNA-guided endonuclease

中图分类号:

刘志国. CRISPR/Cas9系统介导基因组编辑的研究进展[J]. 畜牧兽医学报, 2014, 45(10): 1567-1583.

LIU Zhi-guo. Research Progress on CRISPR/Cas9 Mediated Genome Editing[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(10): 1567-1583.

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