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遗传纠使哺乳动物肠道中的超稳定生物封闭成为可能

Gary W Foo¹, Aathavan S Uruthirapathy¹, Claire Q Zhang¹

¹Department of Biochemistry, Schulich School of Medicine and Dentistry, Western University, London, Ontario N6A 5C1, Canada.

ACS synthetic biology

|September 7, 2025

概括

工程化益生菌面临遗传不稳定性,但STALEMATE (序列结合多层基因基因缓冲环) 提供了一种双层生物封闭解决方案. 这一策略显著降低了哺乳动物肠道中的突变逃脱率.

科学领域:

合成生物学合成生物学
微生物工程微生物工程
基因工程是一种基因工程.

背景情况:

肠道微生物组的不平衡与各种健康状况有关.
工程化益生菌显示出治疗性传递的希望,但遭受遗传不稳定性和突变逃脱.
有效的生物封闭对于工程微生物的安全应用至关重要.

研究的目的:

为工程微生物开发一种新的双层安全生物封闭战略.
为了增强遗传稳定性,并尽量减少突变逃脱率 in vivo.
创建一个强大的系统,用于哺乳动物肠道的治疗应用.

主要方法:

开发了STALEMATE (Sequence enTAngLEd Multi lAyered geneTic buffEring),这是一个纠遗传序列的系统,以创造伪实质性.
叠加的读取框架的素E9免疫蛋白 (Im9) 和热调节的巨核酶 (TSM) 双层无活化.
优化基因表达和序列元素以防止突变和插入.

主要成果:

实现突变逃脱率低于10^-10,与非纠系统的10^-5相比显著改善.
在小鼠胃肠道内,在E. coli Nissle 1917中表现出稳定的等离子体含量超过一周.
在细菌分泌时观察到几乎无法检测的逃逸率.

结论:

STALEMATE为工程细菌提供了一个模块化和有效的生物控制解决方案.
该系统缓冲对突变的无活化,而不需要工程细菌或外源性连接体.
这一战略提高了益生菌用于肠道相关疗法的安全性和可靠性.

关键词:

生物封闭是一种生物封闭.巨大的净化工厂突变逃脱是一种突变逃脱.序列纠是指一个序列纠.在温度调节的内部.毒素 - 抗毒素毒素 - 抗毒素

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