美国麻省理工学院Christopher A. Voigt团队的一项最新研究,实现对肠道共生的多形拟杆菌(Bacteroides thetaiotaomicron)的遗传回路设计自动化。该研究于2020年3月30日在线发表于《自然—生物技术》。
研究人员报道了多形拟杆菌的Cello回路设计自动化软件。首先,他们基于单个向导RNA(CRISPR–dCas9)表征一组基因组整合的NOT/NOR门控。然后,他们将逻辑回路设计为集成对胆汁酸和无水四环素(aTc)做出响应的传感器,其中包括一种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。研究人员发现该回路在实验室条件下至少稳定12天,并在体外人肠模型系统中与原发结肠上皮单层相关细菌一起工作。
Title: Genetic circuit design automation for the gut resident species Bacteroides thetaiotaomicron
Author: Mao Taketani, Jianbo Zhang, Shuyi Zhang, Alexander J. Triassi, Yu-Ja Huang, Linda G. Griffith, Christopher A. Voigt
Issue&Volume: 2020-03-30
Abstract: Bacteroides thetaiotaomicron is a human-associated bacterium that holds promise for delivery of therapies in the gut microbiome1. 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. thetaiotaomicron2,3,4,5. We report implementation of Cello circuit design automation software6 for this species. First, we characterize a set of genome-integrated NOT/NOR gates based on single guide RNAs (CRISPR–dCas9) 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 to function in bacteria associated with a primary colonic epithelial monolayer in an in vitro human gut model system.
DOI: 10.1038/s41587-020-0468-5
Source: https://www.nature.com/articles/s41587-020-0468-5
Nature Biotechnology:《自然—生物技术》,创刊于1996年。隶属于施普林格·自然出版集团,最新IF:31.864
官方网址:https://www.nature.com/nbt/
投稿链接:https://mts-nbt.nature.com/cgi-bin/main.plex