用于自诱导蛋白表达和CRISPRi介导的动态代谢调节的素反应生物传感器工具箱的计算设计
Qianzhen Dong1,2,3, Peng Chen1,2, Zhiru Guo1,3
1Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
Nature communications
|December 22, 2025
概括
我们通过计算设计了一种新的粉生物传感器,使灵敏度增加了20倍. 这使得合成生物学中的新应用成为可能,包括自诱导蛋白表达和对代谢调节的CRISPR干扰.
科学领域:
- 合成生物学 合成生物学
- 蛋白质工程是指蛋白质工程.
- 代谢工程是代谢工程.
背景情况:
- 转录因子 (TF) 生物传感器在合成生物学中至关重要.
- 由于复杂的多域结构,TFs的计算设计具有挑战性.
研究的目的:
- 通过计算设计一种具有增强灵敏度的素反应型TF生物传感器.
- 开发这个生物传感器在生物技术中的新型应用.
主要方法:
- 一种对素敏感的PsiR蛋白的结构指导计算设计.
- 对于PsiR-素生物传感器 (PAB) 的灵敏度和动态范围的表征.
- 在自诱导蛋白表达系统和CRISPR干扰电路中应用PAB.
主要成果:
- 实现了生物传感器灵敏度的20倍增加,将EC50从16mM降低到0.8mM.
- 开发了一种具有10μM至100μM检测范围的PAB.
- 在LacI-IPTG生物传感器中表现出增强的灵敏度.
- 建立了一个无诱导剂的系统,用于自诱导蛋白质表达.
- 实施了一种由素触发的CRISPR干扰电路来调节代谢,将素标位增加了68%,并实现了0.43g/g的葡萄糖产量.
结论:
- 该研究介绍了一种多功能TF工具箱,用于生物技术中素触发调节.
- 开发的生物传感器和系统为代谢工程应用提供了精确的控制.
- 计算设计策略可以克服合成生物学TF工程中的挑战.
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