《生命科学》 2017, 29(6): 610-617

CRISPR-Cas9技术在构建基因线路中的应用

谢海标1，2，刘宇辰1，黄卫人1*

(1 深圳大学第一附属医院-深圳市第二人民医院医学合成生物学临床应用关键技术国家地方
    联合工程实验室，深圳 518036；2 汕头大学医学院临床医学系，汕头 515041)

摘　要：摘　要：随着合成生物学的发展，基因线路在临床医学、生物制剂和化学品生产等多个领域展现出巨大的应用潜力。既往在构建基因线路的过程中往往面临着一个难题——缺乏一个有效的、可编辑的、靶向性高的转录调控因子。原核规律性重复短回文序列簇(CRISPR) 基因编辑/ 修饰系统—— 一种由向导RNA 诱导Cas9 蛋白靶向目的基因的基因编辑工具，具有高效、简单、可编辑等特性，契合了构建基因线路的需要，近来被广泛应用。现对合成基因线路的发展历程以及CRISPR 系统如何改造构建基因线路作一回顾，同时讨论了CRISPR 介导的基因线路的最新进展、潜能和存在的挑战。

The application of CRISPR-Cas9 tools in gene circuits construction

XIE Hai-Biao1，2， LIU Yu-Chen1， HUANG Wei-Ren1*

(1 Department of Urology， State and Local Government Joint Engineering Laboratory of Synthetic Biology Medicine and Clinical Application of Key Technologies， Shenzhen Second Hospital， The First Affiliated Hospital of Shenzhen University， Shenzhen 518036， China; 2 Departments of Clinical Medicine， Shantou University Medical College， Shantou 515041， China)

Abstract: Abstract: With the development of synthetic biology， synthetic gene circuits have showed great applied potentialities in medicine， biology and commodity chemicals. A fundamental challenge in building gene circuits is the shortage of effective， programmable and sequence-specific synthetic transcription factors (TFs). Clustered regularly interspaced short palindromic repeat (CRISPR) system， a CRISPR-associated RNA-guided endonuclease Cas9 targeted genome editing tool， recently has been applied in engineering gene circuits for its pretty unique properties-operability， high efficiency and programmability. We review the evolvement of synthetic gene circuits and how CRISPR system can be used to build it， and discuss recent advances， prospective， underlying challenge in CRISPR-based gene circuits.