Sukegawa Satoru, Toki Seiichi, Saika Hiroaki
Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, 3-1-3 Kannondai, Tsukuba, Ibaraki, 305-8604, Japan.
Graduate School of Nanobioscience, Yokohama City University, Yokohama, Kanagawa, Japan.
Rice (N Y). 2022 Apr 2;15(1):21. doi: 10.1186/s12284-022-00566-4.
Genome editing technology can be used for gene engineering in many organisms. A target metabolite can be fortified by the knockout and modification of target genes encoding enzymes involved in catabolic and biosynthesis pathways, respectively, via genome editing technology. Genome editing is also applied to genes encoding proteins other than enzymes, such as chaperones and transporters. There are many reports of such metabolic engineering using genome editing technology in rice. Genome editing is used not only for site-directed mutagenesis such as the substitution of a single base in a target gene but also for random mutagenesis at a targeted region. The latter enables the creation of novel genetic alleles in a target gene. Recently, genome editing technology has been applied to random mutagenesis in a targeted gene and its promoter region in rice, enabling the screening of plants with a desirable trait from these mutants. Moreover, the expression level of a target gene can be artificially regulated by a combination of genome editing tools such as catalytically inactivated Cas protein with transcription activator or repressor. This approach could be useful for metabolic engineering, although expression cassettes for inactivated Cas fused to a transcriptional activator or repressor should be stably transformed into the rice genome. Thus, the rapid development of genome editing technology has been expanding the scope of molecular breeding including metabolic engineering. In this paper, we review the current status of genome editing technology and its application to metabolic engineering in rice.
基因组编辑技术可用于多种生物体的基因工程。通过基因组编辑技术，分别对编码参与分解代谢和生物合成途径的酶的靶基因进行敲除和修饰，可强化目标代谢产物。基因组编辑还应用于编码酶以外蛋白质的基因，如伴侣蛋白和转运蛋白。有许多关于在水稻中使用基因组编辑技术进行这种代谢工程的报道。基因组编辑不仅用于定点诱变，如在靶基因中替换单个碱基，还用于在靶向区域进行随机诱变。后者能够在靶基因中创建新的遗传等位基因。最近，基因组编辑技术已应用于水稻中靶向基因及其启动子区域的随机诱变，从而能够从这些突变体中筛选出具有理想性状的植株。此外，通过将催化失活的Cas蛋白与转录激活剂或阻遏物等基因组编辑工具组合使用，可以人为调节靶基因的表达水平。尽管与转录激活剂或阻遏物融合的失活Cas的表达盒应稳定转化到水稻基因组中，但这种方法可能对代谢工程有用。因此，基因组编辑技术的快速发展一直在扩大包括代谢工程在内的分子育种范围。在本文中，我们综述了基因组编辑技术的现状及其在水稻代谢工程中的应用。
