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为Corynebacterium stationis开发CRISPR/Cas9基因组编辑工具箱,并将其应用于低素生物合成

Zhilin Ouyang^1,2, Xinyu Zhang^1,2, Xinyi Hou^1,2

¹Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, Guangzhou, 510006, PR China.

Synthetic and systems biotechnology

|July 21, 2025

概括

研究人员为*Corynebacterium stationis*开发了一个CRISPR/Cas9基因组编辑平台,增强了其工业潜力. 这种基因工具箱使生物化学生产能够有效地进行基因编辑和代谢工程.

科学领域:

合成生物学合成生物学
微生物生物技术微生物生物技术
基因组工程是基因组工程.

背景情况:

*Corynebacterium stationis*是一种高GC的阳性菌,具有重要的工业前景.
遗传工具的有限性阻碍了C. stationis的工业应用.
开发高效的基因组编辑系统对于菌株改进至关重要.

研究的目的:

为*C. stationis*建立一个基于CRISPR/Cas9的基因组编辑平台.
为了优化这种细菌的遗传操纵技术.
为了工程 *C. stationis* 增强生产有价值的化合物.

主要方法:

为*C. stationis*优化了电穿孔协议.
构建了用于异质基因表达的功能穿载体.
设计了一个严格规范的CRISPR/Cas9表达系统.
利用CRISPR/Cas9辅助的ssDNA重组进行精确的基因修饰.

主要成果:

使用CRISPR/Cas9系统实现了高删除 (81.2-98.6%) 和插入 (27.5-65.2%) 的效率.
通过ssDNA重组证明了单/三核酸变化>90%的效率.
工程设计的 *C. stationis* 用于低素生物合成,产生0.047g/L.

结论:

为*C. stationis*建立了一个强大的多功能CRISPR/Cas9基因平台.
这个平台显著提高了基因操纵和代谢工程的潜力.
开发的工具将加速C. stationis的工业应用,用于生产生物化学品和生物燃料.

关键词:

这就是CRISPR/Cas9的作用.科里尼细菌已经停滞不前.基因组编辑基因组编辑低桑丁 (Hypoxanthine) 是一种抗氧化剂.转型优化转型优化

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