精准调控CRISPR/Cas9基因编辑技术研究进展

doi:10.16288/j.yczz.20-069

遗传 ›› 2020, Vol. 42 ›› Issue (12): 1168-1177.doi: 10.16288/j.yczz.20-069

精准调控CRISPR/Cas9基因编辑技术研究进展

曹俊霞¹, 王友亮², 王征旭^1,³()

^1. 解放军总医院第七医学中心生物治疗中心,北京 100700
^2. 军事科学院军事医学研究院生物工程研究所,北京 100071
^3. 解放军总医院第一医学中心肝胆胰外科学部,北京 100853

收稿日期:2020-03-13 修回日期:2020-11-02 出版日期:2020-12-17 发布日期:2020-11-25
通讯作者: 王征旭 E-mail:zhxuwang@qq.com
作者简介:曹俊霞,博士,助理研究员,研究方向：生物治疗。E-mail: jxcao2080@139.com
基金资助:
“十三五”科研计划项目资助编号(19LBJ1029C)

Advances in precise regulation of CRISPR/Cas9 gene editing technology

Junxia Cao¹, Youliang Wang², Zhengxu Wang^1,³()

^1. Biotherapy Center, the Seventh Medical Center of PLA General Hospital, Beijing 100700, China
^2. Beijing Institute of Biotechnology, 20 Dongdajie, Beijing 100071, China
^3. The Faculty of Hepato-Pancreato-Biliary Surgery, the First Medical Center of Chinese PLA General Hospital, Beijing 100853, China

Received:2020-03-13 Revised:2020-11-02 Online:2020-12-17 Published:2020-11-25
Contact: Wang Zhengxu E-mail:zhxuwang@qq.com
Supported by:
Supported by the 13th Five Year Research Project No(19LBJ1029C)

摘要/Abstract

摘要：

基因编辑(gene editing)是一种能对细胞和生物体基因组一小段DNA进行定点修饰或者删除、插入的基因工程技术。基因编辑技术在疾病治疗、基因功能调控、基因检测、药物研发和作物育种等方面具有广阔的应用前景,但在应用中也逐渐显现出脱靶、基因毒性等副作用问题。CRISPR (clustered regularly interspaced short palindromic repeats)系统中核酸酶Cas9蛋白能通过与gRNA (guide RNA)结合特异性识别靶DNA并进行酶切反应,由于Cas9蛋白和gRNA在其自身活性、识别位点及结合能力等方面具有不同的特性,因此在应用中可以通过对Cas9蛋白酶的活性以及与靶DNA在时间和空间上的结合进行精准调控,主要调控方法包括使用光、温度和药物等调节Cas9融合蛋白、抗CRISPR蛋白、核酸类和小分子类化合物抑制剂的使用等,从而能有效地防范基因编辑技术的风险和增强精准调控基因编辑技术的实际应用性。本文就目前如何精准调控基因编辑技术,尤其是精准调控CRISPR/Cas9基因编辑技术的方法进行了综述,以期为人类疾病治疗、作物育种、家畜遗传改良和防范生物技术缪用等提供借鉴和研究思路。

关键词: 基因编辑, 基因编辑调控, 抗CRISPR蛋白, 小分子抑制剂

Abstract:

Gene editing is a genetic engineering technology that can modify, delete, or insert a small piece of DNA at a specific point in the genome of cells and organisms. Gene editing technology holds great promises in the fields of disease treatment, gene function regulation, gene detection, drug research and development, and crop breeding. However, side effects, such as off-target editing, genotoxicity and other issues, have gradually emerged in the application. In the CRISPR (clustered regularly interspaced short palindromic repeats) system, the Cas9 nuclease can specifically recognize the target DNA by the base pairing of a guide RNA (gRNA) with the target DNA. Upon target recognition, the two DNA strands are cleaved by distinct domains of the Cas9 nuclease. Since both Cas9 nuclease and gRNA possess different characteristics in their own activities, recognition sites and binding ability to specific target, it is essential to precisely regulate the activity of Cas9 nuclease and gRNA in both time and space manners, thus preventing the risk of side effects and enhancing the precise regulation of the CRISPR/Cas9 gene editing technology. In this review, we summarize the advances in the precise control of gene editing, especially CRISPR/cas9 over several dimensions using fusion Cas9 proteins regulated by light, temperature and drugs, exploiting and screening anti-CRISPRs proteins, synthesizing and identifying small molecules- inhibitors, and developing other therapeutic agents, thereby providing a reference and research ideas for human disease treatment, crop and livestock improvement and prevention of biotechnology misuse.

Key words: gene editing, control of gene editing, anti-CRISPR protein, small molecular inhibitors

曹俊霞, 王友亮, 王征旭. 精准调控CRISPR/Cas9基因编辑技术研究进展[J]. 遗传, 2020, 42(12): 1168-1177.

Junxia Cao, Youliang Wang, Zhengxu Wang. Advances in precise regulation of CRISPR/Cas9 gene editing technology[J]. Hereditas(Beijing), 2020, 42(12): 1168-1177.

图/表 1

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精准调控CRISPR/Cas9基因编辑技术研究进展

Advances in precise regulation of CRISPR/Cas9 gene editing technology

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