Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Tsukuba, Ibaraki, Japan.
Graduate School of Nanobioscience, Yokohama City University, Yokohama, Kanagawa, Japan.
Plant J. 2021 Jun;106(5):1208-1218. doi: 10.1111/tpj.15233. Epub 2021 Apr 24.
Genome-editing technologies consisting of targeted mutagenesis and gene targeting enable us to modify genes of interest rapidly and precisely. The discovery in 2012 of CRISPR/Cas9 systems and their development as sequence-specific nucleases has brought about a paradigm shift in biology. Initially, CRISPR/Cas9 was applied in targeted mutagenesis to knock out a target gene. Thereafter, advances in genome-editing technologies using CRISPR/Cas9 developed rapidly, with base editing systems for transition substitution using a combination of Cas9 nickase and either cytidine or adenosine deaminase being reported in 2016 and 2017, respectively, and later in 2021 bringing reports of transversion substitution using Cas9 nickase, cytidine deaminase and uracil DNA glycosylase. Moreover, technologies for gene targeting and prime editing systems using DNA or RNA as donors have also been developed in recent years. Besides these precise genome-editing strategies, reports of successful chromosome engineering using CRISPR/Cas9 have been published recently. The application of genome editing to crop breeding has advanced in parallel with the development of these technologies. Genome-editing enzymes can be introduced into plant cells, and there are now many examples of crop breeding using genome-editing technologies. At present, it is no exaggeration to say that we are now in a position to be able to modify a gene precisely and rearrange genomes and chromosomes in a predicted way. In this review, we introduce and discuss recent highlights in the field of precise gene editing, chromosome engineering and genome engineering technology in plants.
基因组编辑技术包括靶向诱变和基因靶向，使我们能够快速、精确地修饰感兴趣的基因。2012 年 CRISPR/Cas9 系统的发现及其作为序列特异性核酸酶的发展，带来了生物学的范式转变。最初，CRISPR/Cas9 被应用于靶向诱变以敲除靶基因。此后，利用 CRISPR/Cas9 的基因组编辑技术迅速发展，分别于 2016 年和 2017 年报道了使用 Cas9 切口酶和胞嘧啶或腺嘌呤脱氨酶的碱基编辑系统进行转换替换，随后在 2021 年报道了使用 Cas9 切口酶、胞嘧啶脱氨酶和尿嘧啶 DNA 糖基化酶进行颠换替换。此外，近年来还开发了用于基因靶向和使用 DNA 或 RNA 作为供体的 Prime 编辑系统。除了这些精确的基因组编辑策略外，最近还报道了使用 CRISPR/Cas9 成功进行染色体工程的报告。基因组编辑在作物育种中的应用与这些技术的发展并行推进。基因组编辑酶可以被引入植物细胞，现在有许多利用基因组编辑技术进行作物育种的例子。目前，可以毫不夸张地说，我们现在能够以精确的方式修饰基因，并以预测的方式重新排列基因组和染色体。在这篇综述中，我们介绍和讨论了植物中精确基因编辑、染色体工程和基因组工程技术的最新亮点。
