The CRISPR-based gene editing technology has become more and more powerful in genome manipulation for agricultural breeding, with numerous improved toolsets springing up. In recent years, many CRISPR toolsets for gene editing, such as base editors (BEs), CRISPR interference (CRISPRi), CRISPR activation (CRISPRa), and plant epigenetic editors (PEEs), have been developed to clarify gene function and full-level gene regulation. Here, we comprehensively summarize the application and capacity of the different CRISPR toolsets in the study of plant gene expression regulation, highlighting their potential application in gene regulatory networks' analysis. The general problems in CRISPR application and the optimal solutions in the existing schemes for high-throughput gene function analysis are also discussed. The CRISPR toolsets targeting gene manipulation discussed here provide new solutions for further genetic improvement and molecular breeding of crops.

 摘 要 

基于CRISPR的基因编辑技术在农业育种的基因组操作中应用得越来越多，涌现出许多经过改良的工具包。近年来，研究人员已经开发出了许多用于基因编辑的CRISPR工具包，如碱基编辑器（BEs）、CRISPR干扰（CRISPRi）、CRISPR激活（CRISPRa）以及植物表观遗传学编辑器（PEEs），用来阐明基因功能和各种水平上的基因调控机制。本文中，作者全面总结了不同CRISPR工具包在植物基因表达调控研究中的应用和能力，重点介绍了其在基因调控网络分析中的潜在应用。作者还讨论了CRISPR应用中的共性问题，以及现在高通量基因功能分析方案中的最优解决方案。本文所讨论的以基因操纵为目标的CRISPR工具包为作物的进一步遗传改良和分子育种提供了新的解决方案。

 陈坤明