Graduate School of Technology, Industrial and Social Sciences, Tokushima University, Tokushima, Japan.
BMC Plant Biol. 2020 May 25;20(1):234. doi: 10.1186/s12870-020-02385-5.
Traditionally, generation of new plants with improved or desirable features has relied on laborious and time-consuming breeding techniques. Genome-editing technologies have led to a new era of genome engineering, enabling an effective, precise, and rapid engineering of the plant genomes. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) has emerged as a new genome-editing tool, extensively applied in various organisms, including plants. The use of CRISPR/Cas9 allows generating transgene-free genome-edited plants ("null segregants") in a short period of time. In this review, we provide a critical overview of the recent advances in CRISPR/Cas9 derived technologies for inducing mutations at target sites in the genome and controlling the expression of target genes. We highlight the major breakthroughs in applying CRISPR/Cas9 to plant engineering, and challenges toward the production of null segregants. We also provide an update on the efforts of engineering Cas9 proteins, newly discovered Cas9 variants, and novel CRISPR/Cas systems for use in plants. The application of CRISPR/Cas9 and related technologies in plant engineering will not only facilitate molecular breeding of crop plants but also accelerate progress in basic research.
传统上，具有改良或理想特征的新植物的产生依赖于费力且耗时的繁殖技术。基因组编辑技术开创了基因组工程的新纪元，使植物基因组的有效、精确和快速工程成为可能。成簇规律间隔短回文重复 (CRISPR)/CRISPR 相关蛋白 9 (CRISPR/Cas9) 已成为一种新的基因组编辑工具，广泛应用于包括植物在内的各种生物体。CRISPR/Cas9 的使用允许在短时间内产生无转基因基因组编辑植物（“纯合子”）。在这篇综述中，我们批判性地概述了 CRISPR/Cas9 衍生技术在诱导基因组靶位点突变和控制靶基因表达方面的最新进展。我们强调了将 CRISPR/Cas9 应用于植物工程的主要突破，以及生产纯合子的挑战。我们还提供了关于工程 Cas9 蛋白、新发现的 Cas9 变体以及用于植物的新型 CRISPR/Cas 系统的最新进展。CRISPR/Cas9 及相关技术在植物工程中的应用不仅将促进作物的分子育种，也将加速基础研究的进展。
