Genetic circuit design automation for the gut resident species Bacteroides thetaiotaomicron.

Genetic circuit design automation for the gut resident species Bacteroides thetaiotaomicron.
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肠道居民物种的遗传回路设计自动化菌虫菌属。

DOI:
10.1038/s41587-020-0468-5
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发表时间:
2020-08
影响因子:
46.9
通讯作者:
Voigt CA
Voigt CA
中科院分区:
工程技术1区
文献类型:
--
作者:
Taketani M;Zhang J;Zhang S;Triassi AJ;Huang YJ;Griffith LG;Voigt CA

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多形拟杆菌是一种人类肠道细菌,有望在肠道微生物组中提供治疗。治疗性细菌将受益于响应肠道内外条件而开启不同基因表达程序的能力;然而,在B中,将它们联合收割机结合的调节部分和方法的可用性受到限制。太多了。我们报告实现大提琴电路设计自动化软件,为这个物种。首先,我们描述了一组基于sgRNA(CRISPR/Cas9)的基因组整合的NOT/NOR门,以通知Cello的Bt用户约束文件(UCF)。然后,逻辑电路被设计成集成对胆汁酸和脱水四环素(aTc)做出响应的传感器,包括用于区分与生物生产、人类肠道和释放后相关的环境的传感器。发现该回路在实验室条件下稳定至少12天,并且在体外人肠道模型系统中与原代结肠上皮单层相关的细菌中起作用。
Bacteroides thetaiotaomicron is a human gut bacterium that holds promise for delivery of therapies in the gut microbiome. Therapeutic bacteria would benefit from the ability to turn on different programs of gene expression in response to conditions inside and outside of the gut; however, the availability of regulatory parts and methods to combine them have been limited in B. thetaiotaomicron. We report implementation of Cello circuit design automation software for this species. First, we characterize a set of genome-integrated NOT/NOR gates based on sgRNAs (CRISPR/Cas9) to inform a Bt user constraint file (UCF) for Cello. Then, logic circuits are designed to integrate sensors that respond to bile-acid and anhydrotetracycline (aTc), including one created to distinguish between environments associated with bioproduction, the human gut, and after release. This circuit was found to be stable under laboratory conditions for at least 12 days and function in bacteria associated with a primary colonic epithelial monolayer in an in vitro human gut model system.
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发表时间: 2016-01
期刊: Nature reviews. Microbiology
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