Zhou Junming, Luan Xinchao, Liu Yixuan, Wang Lixue, Wang Jiaxin, Yang Songnan, Liu Shuying, Zhang Jun, Liu Huijing, Yao Dan
College of Life Sciences, Jilin Agricultural University, Changchun 130118, China.
College of Agronomy, Jilin Agricultural University, Changchun 130118, China.
Plants (Basel). 2023 Mar 28;12(7):1478. doi: 10.3390/plants12071478.
Following recent developments and refinement, CRISPR-Cas9 gene-editing technology has become increasingly mature and is being widely used for crop improvement. The application of CRISPR/Cas9 enables the generation of transgene-free genome-edited plants in a short period and has the advantages of simplicity, high efficiency, high specificity, and low production costs, which greatly facilitate the study of gene functions. In plant molecular breeding, the gene-editing efficiency of the CRISPR-Cas9 system has proven to be a key step in influencing the effectiveness of molecular breeding, with improvements in gene-editing efficiency recently becoming a focus of reported scientific research. This review details strategies and methods for improving the efficiency of CRISPR/Cas9 gene editing in plant molecular breeding, including Cas9 variant enzyme engineering, the effect of multiple promoter driven Cas9, and gRNA efficient optimization and expression strategies. It also briefly introduces the optimization strategies of the CRISPR/Cas12a system and the application of BE and PE precision editing. These strategies are beneficial for the further development and optimization of gene editing systems in the field of plant molecular breeding.
随着近期的发展和完善,CRISPR-Cas9基因编辑技术日益成熟,并被广泛应用于作物改良。CRISPR/Cas9的应用能够在短时间内产生无转基因的基因组编辑植物,具有操作简单、效率高、特异性强和生产成本低等优点,极大地促进了基因功能的研究。在植物分子育种中,CRISPR-Cas9系统的基因编辑效率已被证明是影响分子育种效果的关键步骤,提高基因编辑效率近来成为报道科研的重点。本文综述了提高植物分子育种中CRISPR/Cas9基因编辑效率的策略和方法,包括Cas9变体酶工程、多个启动子驱动Cas9的效果以及gRNA高效优化和表达策略。还简要介绍了CRISPR/Cas12a系统的优化策略以及碱基编辑器(BE)和引导编辑(PE)精准编辑的应用。这些策略有利于植物分子育种领域基因编辑系统的进一步发展和优化。