In vitro Cas9-assisted editing of modular polyketide synthase genes to produce desired natural product derivatives

Kei Kudo, Takuya Hashimoto, Junko Hashimoto, Ikuko Kozone, Noritaka Kagaya, Reiko Ueoka, Takehiro Nishimura, Mamoru Komatsu, Hikaru Suenaga, Haruo Ikeda, Kazuo Shin-ya

研究成果: Article査読

22 被引用数 (Scopus)

抄録

One major bottleneck in natural product drug development is derivatization, which is pivotal for fine tuning lead compounds. A promising solution is modifying the biosynthetic machineries of middle molecules such as macrolides. Although intense studies have established various methodologies for protein engineering of type I modular polyketide synthase(s) (PKSs), the accurate targeting of desired regions in the PKS gene is still challenging due to the high sequence similarity between its modules. Here, we report an innovative technique that adapts in vitro Cas9 reaction and Gibson assembly to edit a target region of the type I modular PKS gene. Proof-of-concept experiments using rapamycin PKS as a template show that heterologous expression of edited biosynthetic gene clusters produced almost all the desired derivatives. Our results are consistent with the promiscuity of modular PKS and thus, our technique will provide a platform to generate rationally designed natural product derivatives for future drug development.

本文言語English
論文番号4022
ジャーナルNature communications
11
1
DOI
出版ステータスPublished - 2020 12月 1
外部発表はい

ASJC Scopus subject areas

  • 化学一般
  • 生化学、遺伝学、分子生物学一般
  • 物理学および天文学一般

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