Rebooting Synthetic Phage-Inducible Chromosomal Islands: One Method to Forge Them All.
Rebooting Synthetic Phage-Inducible Chromosomal Islands: One Method to Forge Them All.
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DOI：
10.34133/2020/5783064
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发表时间：
2020
影响因子：
--
通讯作者：
Penadés JR
中科院分区：
文献类型：
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作者：
Ibarra-Chávez R;Haag AF;Dorado-Morales P;Lasa I;Penadés JR
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.
