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工程化的细胞因子基编辑器,使得Streptomyces中的广泛和高可靠性基因编辑成为可能

Jian Wang¹, Ke Wang¹, Zhe Deng¹

¹Department of Hematology, Zhongnan Hospital of Wuhan University, School of Pharmaceutical Sciences, Wuhan University, Wuhan, 430071, China.

Nature communications

|July 6, 2024

概括

一个新的基准编辑器,eSCBE3-NG-Hypa,有效地修改Streptomyces基因组,克服了诸如高GC含量等挑战. 这种工具通过非激活竞争的生物合成基因集群来增强阿弗梅克丁的产生.

科学领域:

微生物遗传学和合成生物学
抗生素生物合成和代谢工程

背景情况:

基础编辑 (BE) 工具面临的局限性,如原空间隔离图案 (PAM) 约束和非目标效应.
对于抗生素生产至关重要的Streptomyces物种,由于高GC含量和复杂的基因组,存在独特的挑战.

研究的目的:

为Streptomyces开发一个高效和特定的基础编辑器.
为了改造Streptomyces avermitilis以提高有价值化合物的生产.

主要方法:

开发eSCBE3-NG-Hypa,这是一个新的基础编辑器,可以识别NG PAM.
在Streptomyces avermitilis中应用eSCBE3-NG-Hypa进行精确的基因编辑.
在多基合成酶 (PKS) 和竞争基因集群中脱水酶域的非激活.

主要成果:

eSCBE3-NG-Hypa表现出高活力和特异性,即使在富含GC的地点.
精确无活化阿维美克生物合成基因集群 (BGCs) 导致阿维美克B1a产量增加 (4.45倍).
工程突变将产量从阿弗梅克丁转移到奥利戈米辛,揭示了BGC竞争.

结论:

eSCBE3-NG-Hypa是精确基因组工程在Streptomyces的一个强大的工具.
这项技术为优化天然产品生物合成的代谢工程策略提供了便利.
了解BGC竞争有助于最大限度地提高所需化合物的产量.

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