Abdelrahman Mohamed, Wei Zheng, Rohila Jai S, Zhao Kaijun
National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing, China.
Rice Research and Training Center, Field Crops Research Institute, Agricultural Research Center, Kafr El-Shaikh, Egypt.
Front Plant Sci. 2021 Oct 6;12:721203. doi: 10.3389/fpls.2021.721203. eCollection 2021.
Multiplex genome-editing (MGE) technologies are recently developed versatile bioengineering tools for modifying two or more specific DNA loci in a genome with high precision. These genome-editing tools have greatly increased the feasibility of introducing desired changes at multiple nucleotide levels into a target genome. In particular, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) [CRISPR/Cas] system-based MGE tools allow the simultaneous generation of direct mutations precisely at multiple loci in a gene or multiple genes. MGE is enhancing the field of plant molecular biology and providing capabilities for revolutionizing modern crop-breeding methods as it was virtually impossible to edit genomes so precisely at the single base-pair level with prior genome-editing tools, such as zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs). Recently, researchers have not only started using MGE tools to advance genome-editing applications in certain plant science fields but also have attempted to decipher and answer basic questions related to plant biology. In this review, we discuss the current progress that has been made toward the development and utilization of MGE tools with an emphasis on the improvements in plant biology after the discovery of CRISPR/Cas9. Furthermore, the most recent advancements involving CRISPR/Cas applications for editing multiple loci or genes are described. Finally, insights into the strengths and importance of MGE technology in advancing crop-improvement programs are presented.
多重基因组编辑（MGE）技术是最近开发的多功能生物工程工具，用于在基因组中高精度地修饰两个或更多特定的DNA位点。这些基因组编辑工具极大地提高了在多个核苷酸水平上向目标基因组引入所需变化的可行性。特别是，基于成簇规律间隔短回文重复序列（CRISPR）/CRISPR相关蛋白（Cas）[CRISPR/Cas]系统的MGE工具能够在一个基因或多个基因的多个位点同时精确产生直接突变。MGE正在推动植物分子生物学领域的发展，并为革新现代作物育种方法提供能力，因为使用诸如锌指核酸酶（ZFN）和转录激活因子样效应物核酸酶（TALEN）等先前的基因组编辑工具几乎不可能在单碱基对水平上如此精确地编辑基因组。最近，研究人员不仅开始使用MGE工具来推进某些植物科学领域的基因组编辑应用，还试图解读和回答与植物生物学相关的基本问题。在这篇综述中，我们讨论了MGE工具在开发和利用方面取得的当前进展，重点是CRISPR/Cas9发现后植物生物学的进展。此外，还描述了涉及CRISPR/Cas应用于编辑多个位点或基因的最新进展。最后，阐述了MGE技术在推进作物改良计划中的优势和重要性。
