CRISPR⁃Cas9技术在植物次生代谢物生产中的研究进展

doi:10.19586/j.2095-2341.2022.0056

生物技术进展 ›› 2022, Vol. 12 ›› Issue (6): 806-816.DOI: 10.19586/j.2095-2341.2022.0056

CRISPR⁃Cas9技术在植物次生代谢物生产中的研究进展

杨洋(), 王凤林, 刘德, 罗园园, 朱建华()

暨南大学药学院，广州 510632

收稿日期:2022-04-14 接受日期:2022-08-30 出版日期:2022-11-25 发布日期:2022-11-30
通讯作者: 朱建华
作者简介:杨洋 E-mail: yang3317@qq.com；
基金资助:
国家自然科学基金项目(81673571);广东省自然科学杰出青年基金(2016A030306009);广州市基础研究项目(202201010107);广州市市校联合资助基础研究项目(SL2023A03J00675)

Research Progress of CRISPR‍⁃‍Cas9 Technology on the Production of Plant Secondary Metabolites

Yang YANG(), Fenglin WANG, De LIU, Yuanyuan LUO, Jianhua ZHU()

College of Pharmacy，Jinan University，Guangzhou 510632，China

Received:2022-04-14 Accepted:2022-08-30 Online:2022-11-25 Published:2022-11-30
Contact: Jianhua ZHU

摘要/Abstract

摘要：

基因编辑（gene editing）技术可以对目的基因进行定点插入、敲除和置换。基于CRISPR-Cas9的基因编辑技术是继锌指核酸酶和转录激活样效应物核酸酶之后的第3代基因编辑技术。近年来，CRISPR-Cas9系统作为研究的热点被广泛应用于医学、药学、植物学、动物学和微生物学等领域，但其在植物次生代谢物领域的应用还处于探索时期。阐述了基于CRISPR-Cas9基因编辑技术的发展历程、工作原理和几种常用的基因编辑方法及其应用实例，总结了CRISPR-Cas9技术在对植物次生代谢产物研究方面的应用。利用CRISPR-Cas9系统可对植物基因组进行定点敲除、突变和插入，以达到提高植物次生代谢物含量、改良作物品质和提高植物抗性等目的。该技术已在植物次生代谢物生物合成关键酶基因的编辑等方面显示出越来越重要的作用。

关键词: 基因编辑, CRISPR-Cas9, 植物次级代谢物

Abstract:

Gene editing technology can carry out target gene insertion， knockout and replacement. Gene editing technology based on CRISPR-Cas9 was the third generation of gene editing technology after zinc finger nuclease and transcription activator-like effector nuclease. In recent years， CRISPR-Cas9 has been applied in multi-aspects as a research hotspot， such as medicine， pharmacy， botany， zoology and microbiology. However， its application in the field of plant secondary metabolites is still in the exploratory period. The development history of gene editing technologies， the principle of CRISPR-Cas9 and the application of technologies were briefly introduced. More importantly， the application of CRISPR-Cas9 technology on plant secondary metabolites production were summarized. CRISPR-Cas9 system can be used to knock out， mutate and insert the plant genome at fixed points， so as to improve the content of plant secondary metabolites， improve crop quality and plant resistance. This technology has shown an increasingly important role in the edition of key enzyme genes involved in the biosynthesis of plant secondary metabolites.

Key words: gene editing, CRISPR-Cas9, plant secondary metabolites

中图分类号:

Q943.2

杨洋, 王凤林, 刘德, 罗园园, 朱建华. CRISPR⁃Cas9技术在植物次生代谢物生产中的研究进展[J]. 生物技术进展, 2022, 12(6): 806-816.

Yang YANG, Fenglin WANG, De LIU, Yuanyuan LUO, Jianhua ZHU. Research Progress of CRISPR‍⁃‍Cas9 Technology on the Production of Plant Secondary Metabolites[J]. Current Biotechnology, 2022, 12(6): 806-816.

图/表 2

图1 天然CRISPR?Cas9系统[19]A：外源DNA首次入侵细菌，产生crRNA/tracrRNA/Cas9复合物。①—Cas1和Cas2蛋白识别外源DNA的PAM序列；②—识别后剪切DNA临近序列；③—在其他酶的协助下将剪切下来的DNA片段作为间隔序列插入到临近的CRISPR序列中，然后进行DNA修复，修补双链缺口；④—细菌自身附近的CRISPR序列经转录和加工表达crRNA和tracrRNA；⑤—crRNA、tracrRNA和Cas9蛋白组合成为复合物备用。B：相同外源DNA再次入侵细菌后靶基因的识别与切割。①—crRNA/tracrRNA/Cas9复合物识别并精确定位至与crRNA互补的外源DNA的PAM序列；②—crRNA/tracrRNA/Cas9复合物对外源DNA进行精确切割，使外源DNA双链断裂而失活。

Fig. 1 Natural CRISPR?Cas9 system[19]

图2 基于CRISPR?Cas9的基因编辑技术原理[6]

Fig. 2 Principle of gene editing technology based on CRISPR?Cas9[6]

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CRISPR⁃Cas9技术在植物次生代谢物生产中的研究进展

Research Progress of CRISPR‍⁃‍Cas9 Technology on the Production of Plant Secondary Metabolites

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