Igarashi Takayuki, Katayama Takuya, Maruyama Jun-Ichi
Department of Biotechnology, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
Collaborative Research Institute for Innovative Microbiology, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
Biosci Biotechnol Biochem. 2023 Sep 21;87(10):1236-1248. doi: 10.1093/bbb/zbad101.
Aspergillus sojae has traditionally been used in soy sauce brewing. Genetic modification techniques have been established in A. sojae, but it is difficult to apply them to various industrial strains. Although we have previously developed a CRISPR/Cpf1 system for genetic modification of A. sojae, another genome editing system was required for versatile modification. In addition, repetitive genetic modification using the CRISPR system has not been established in A. sojae. In this study, we demonstrated mutagenesis, gene deletion/integration, and large deletion of a chromosomal region in A. sojae using the CRISPR/Cas9 system. We also successfully performed repetitive genetic modification using a method that involved forced recycling of genome-editing plasmids. Moreover, we demonstrated that the effects of genetic modification related to soy sauce brewing differed among A. sojae industrial strains. These results showed that our technique of using the CRISPR/Cas9 system is a powerful tool for genetic modification in A. sojae.
米曲霉传统上用于酱油酿造。米曲霉的基因改造技术已经建立，但将其应用于各种工业菌株却很困难。尽管我们之前已经开发了用于米曲霉基因改造的CRISPR/Cpf1系统，但仍需要另一种基因组编辑系统来进行通用改造。此外，米曲霉中尚未建立使用CRISPR系统的重复基因改造方法。在本研究中，我们展示了利用CRISPR/Cas9系统在米曲霉中进行诱变、基因缺失/整合以及染色体区域的大片段缺失。我们还通过一种涉及强制循环基因组编辑质粒的方法成功进行了重复基因改造。此外，我们证明了与酱油酿造相关的基因改造效果在米曲霉工业菌株中存在差异。这些结果表明，我们使用CRISPR/Cas9系统的技术是米曲霉基因改造的有力工具。
