《生命科学》 2018, 30(9): 980-986

化繁为简——植物多基因编辑体系的优化

王木桂，朱健康*

(中国科学院上海植物逆境生物学研究中心，中国科学院分子植物科学卓越创新中心，上海 201602)

摘　要：摘　要：多基因编辑技术对于基因家族和通路的研究以及作物改良具有十分重要的意义。利用CRISPR-Cas系统在植物中进行多基因编辑需要同时表达Cas 基因和多个sgRNA，组成一个多转录元件系统。为了简化多基因编辑体系，人们利用RNA 自剪切工具和植物内源的RNA 加工机制把多转录元件改造成双转录元件和单转录元件。该文介绍了植物多基因编辑体系由繁至简不断优化过程中的经典案例，同时对存在的问题及发展的方向进行讨论。

Simplification of multiplex gene editing systems in plants

WANG Mu-Gui， ZHU Jian-Kang*

(Shanghai Center for Plant Stress Biology and Center for Excellence in Molecular Plant Sciences，Chinese Academy of Sciences， Shanghai 201602， China)

Abstract: Abstract: Multiplex gene editing is a powerful tool for targeting plant gene families and is of great importance for crop improvement. Generally， the Cas endonuclease is expressed from an RNA polymerase (pol) II promoter， while sgRNA is expressed from a pol III promoter， such as U6 or U3 for gene editing with the CRISPR-Cas system. For multiplex gene editing， multiple sgRNAs are usually driven by separate pol III promoters. To simplify multiplex gene editing in plants， these sgRNAs can be expressed as a single transcript， which generates functional individual sgRNAs after processing by various RNA processing systems. Further， the Cas endonuclease and its sgRNA array can be co-expressed from a single pol II promoter， without any additional RNA processing machinery. In this review， we discuss some examples and potential problems in the optimization of multiplex gene editing systems in plants.