一个由定数感应控制的I型CRISPRi工具包,用于动态调节代谢流量
Han Zheng1, Chengyao Mao1, Siyao Chen1
1College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, Zhejiang 310014, China.
Nucleic acids research
|July 19, 2025
概括
我们为动态代谢工程开发了一种新的定量感应控制的CRISPR干扰 (QICi) 工具包. 这种工具增强了微生物生产有价值的化合物,如d-pantothenic acid和riboflavin.
科学领域:
- 合成生物学 合成生物学
- 代谢工程是代谢工程.
- 微生物生物技术 微生物生物技术
背景情况:
- 使用定量感应 (QS) 和CRISPR系统的动态代谢调节正在推动合成生物学.
- 整合QS与CRISPR进行动态控制尚未得到充分探索.
研究的目的:
- 开发一种QS控制的I型CRISPR干扰 (QICi) 工具包,用于动态基因表达调制.
- 优化QICi组件并实现其用于增强微生物生物合成.
主要方法:
- 开发了一个QS控制的I型CRISPR干扰 (QICi) 工具包.
- 简化了CRISPR RNA (crRNA) 矢量结构和优化了QS组件 (PhroQ和RapQ).
- 在Bacillus subtilis中实施了QICi,用于d-pantothenic acid (DPA) 和 riboflavin (RF) 生物合成.
主要成果:
- 在QICi疗效方面实现了两倍增强.
- 在料批发发酵中,DPA度升至14.97g/l.
- 通过QICi介导的代谢重新连接,RF产量增加了2.49倍.
结论:
- QICi工具包为微生物新陈代谢提供了强大的动态控制.
- 该工具通过增强关键化合物的生产,促进可持续的生物制造.
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