与CRISPRi连接的多模块负反循环解决了获胜者夺取所有资源的竞争
Sadikshya Rijal1, Kylie Standage-Beier1, Rong Zhang1
1School of Biological and Health Systems Engineering, Arizona State University, Tempe, Arizona, 85281, United States.
bioRxiv : the preprint server for biology
|June 4, 2025
概括
合成基因电路经常受到资源竞争的影响,导致获胜者获取全部 (WTA) 行为. 一个使用CRISPR干扰 (CRISPRi) 的新型负竞争性监管 (NCR) 控制器有效地抑制了WTA,增强了合成生物学电路模块化.
科学领域:
- 合成生物学 合成生物学
- 基因工程是一种基因工程.
- 系统生物学 系统生物学
背景情况:
- 细胞资源的限制导致合成基因电路模块之间的意外相互作用,减少模块化.
- 基因回路中的积极反可以加剧资源竞争,导致一个模块占据主导地位的获胜者获取全部 (WTA) 行为.
研究的目的:
- 用CRISPR干扰 (CRISPRi) 来实验实施和评估一种负竞争性监管 (NCR) 控制器.
- 评估NCR控制器在减轻双自激活和级联双可开关合成基因电路中的WTA行为方面的有效性.
主要方法:
- 可调节的dCas9基因的染色体整合.
- 模块特定导向RNA (gRNA) 的设计,用于通过竞争获得有限的dCas9.9进行自我抑制.
- 在双自激活和级联双可位开关电路中实现NCR控制器.
主要成果:
- 与没有dCas9介导抑制的控制电路相比,NCR控制器显著增加了模块的协同激活.
- 通过NCR控制器有效地抑制了WTA行为,促进了平衡的模块激活.
- 量化结果表明,资源从更活跃的模块转移到不那么活跃的模块,实现了成功的重新分配.
结论:
- 负竞争性监管 (NCR) 控制器是合成基因电路中管理资源竞争的有效策略.
- 通过减轻意想不到的相互作用和促进平衡的模块功能,NCR提高了合成基因电路的模块性.
- 基于CRISPRi的NCR提供了一种可调节和强大的方法来提高合成基因电路的性能.
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