Rebooting Synthetic Phage-Inducible Chromosomal Islands: One Method to Forge Them All.

Q2 Agricultural and Biological Sciences
生物设计研究(英文) Pub Date : 2020-05-11 eCollection Date: 2020-01-01 DOI:10.34133/2020/5783064
Rodrigo Ibarra-Chávez, Andreas F Haag, Pedro Dorado-Morales, Iñigo Lasa, José R Penadés
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引用次数: 7

Abstract

Phage-inducible chromosomal islands (PICIs) are a widespread family of mobile genetic elements, which have an important role in bacterial pathogenesis. These elements mobilize among bacterial species at extremely high frequencies, representing an attractive tool for the delivery of synthetic genes. However, tools for their genetic manipulation are limited and timing consuming. Here, we have adapted a synthetic biology approach for rapidly editing of PICIs in Saccharomyces cerevisiae based on their ability to excise and integrate into the bacterial chromosome of their cognate host species. As proof of concept, we engineered several PICIs from Staphylococcus aureus and Escherichia coli and validated this methodology for the study of the biology of these elements by generating multiple and simultaneous mutations in different PICI genes. For biotechnological purposes, we also synthetically constructed PICIs as Trojan horses to deliver different CRISPR-Cas9 systems designed to either cure plasmids or eliminate cells carrying the targeted genes. Our results demonstrate that the strategy developed here can be employed universally to study PICIs and enable new approaches for diagnosis and treatment of bacterial diseases.

Abstract Image

Abstract Image

Abstract Image

重启合成噬菌体诱导的染色体岛:一种锻造它们的方法。
噬菌体诱导型染色体岛(PICIs)是一个广泛分布的可移动遗传元件家族,在细菌发病机制中发挥着重要作用。这些元素在细菌物种中以极高的频率移动,代表了一种有吸引力的合成基因传递工具。然而,用于基因操作的工具是有限的,而且耗时。在这里,我们采用了一种合成生物学方法,根据其切割并整合到同源宿主物种的细菌染色体中的能力,快速编辑酿酒酵母中的PICI。作为概念的证明,我们从金黄色葡萄球菌和大肠杆菌中设计了几种PICI,并通过在不同的PICI基因中产生多个同时突变来验证该方法用于研究这些元素的生物学。出于生物技术目的,我们还合成了PICI作为特洛伊木马,以提供不同的CRISPR-Cas9系统,该系统旨在治愈质粒或消除携带靶基因的细胞。我们的研究结果表明,这里开发的策略可以普遍用于研究PICI,并为细菌性疾病的诊断和治疗提供新的方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
3.90
自引率
0.00%
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审稿时长
12 weeks
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