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基诺Mine:一种基于CRISPR-Cas9的杀死开关,用于生物制Pseudomonas putida

Enrique Asin-Garcia^1,2, Maria Martin-Pascual¹, Claudia de Buck¹

¹Laboratory of Systems and Synthetic Biology, Wageningen University & Research, Wageningen, Netherlands.

Frontiers in bioengineering and biotechnology

|October 1, 2024

概括

我们开发了GenoMine保护,一种用于Pseudomonas putida的CRISPR-Cas9杀死开关,使用基因组元素触发细胞死亡. 该系统提供对微生物生存的可编程控制,用于生物控制和其他应用.

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
基因工程是一种基因工程.

背景情况:

合成遗传电路使微生物细胞活力的可编程控制成为可能.
伪虫 (Pseudomonas putida) 是生物技术中的关键生物体,需要强大的控制机制.

研究的目的:

开发一种基于CRISPR-Cas9的杀死开关,名为GenoMine保护,用于Pseudomonas putida.
研究各种调节机制来控制杀死开关的激活和Cas9的表达.

主要方法:

利用CRISPR-Cas9技术,将重复的基因组元素作为Pseudomonas putida中的目标.
实施诱导表达系统和肋骨调节器来控制Cas9活性及其抑制剂 (AcrIIA4,TetR).
在不同的诱导和抑制条件下评估细胞存活率.

主要成果:

在 Cas9 激活时,GenoMine 保护剂有效抑制了 Pseudomonas putida 细胞存活率.
直接诱导Cas9和抑制其抑制剂都导致了细胞死亡.
通过转录和后转录调节证明了系统的功能.

结论:

基诺Mine保护器提供了一个强大的杀死开关,用于Pseudomonas putida.
该系统的监管灵活性允许在生物封闭之外的各种应用.
通过合成遗传电路,可以实现对细胞活力的可编程控制.

关键词:

在CRISPR-Cas9中介的生物控制中.在P. putida中可以看到.遗传电路是一个遗传电路.杀死开关是一个关键.带调节器是一种带调节器.

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