Rebooting synthetic phage-inducible chromosomal islands: one method to forge them all

Ibarra-Chávez, R., Haag, A. F. , Dorado-Morales, P., Lasa, I. and Penadés, J. R. (2020) Rebooting synthetic phage-inducible chromosomal islands: one method to forge them all. BioDesign Research, 2020, 5783064. (doi: 10.34133/2020/5783064)

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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.

Item Type:Articles
Glasgow Author(s) Enlighten ID:Ibarra Chavez, Mr Rodrigo and Penades, Prof Jose R and Haag, Dr Andreas
Creator Roles:
Ibarra-Chávez, R.Conceptualization, Methodology, Investigation, Writing – original draft
Penadés, J. R.Conceptualization, Methodology, Writing – original draft, Funding acquisition, Supervision
Haag, A. F.Methodology, Investigation
Authors: Ibarra-Chávez, R., Haag, A. F., Dorado-Morales, P., Lasa, I., and Penadés, J. R.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:BioDesign Research
Publisher:American Association for the Advancement of Science
Copyright Holders:Copyright © 2020 Rodrigo Ibarra-Chávez et al.
First Published:First published in BioDesign Research 2020: 5783064
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
170721Molecular biology of the PICIs, a novel and widespread family of mobile genetic elements involved in bacterial virulenceJose R PenadesMedical Research Council (MRC)MR/M003876/1III - Bacteriology - Dr J Penades
304055Deciphering Gram-negative phage-inducible chromosomal island strategies for spreading in natureJose R PenadesMedical Research Council (MRC)MRS00940X/1III - Bacteriology
172242Understanding a novel mechanim involving pathogenity islands in the transfer of unlinked chromosomal virulence genesJose R PenadesBiotechnology and Biological Sciences Research Council (BBSRC)BB/N002873/1Institute of Infection, Immunity & Inflammation
302971Helper and satellite pathogenicity islands: the discovery of two novel subcellular elements with a huge impact on bacterial pathogenesis and evolutionJose R PenadesBiotechnology and Biological Sciences Research Council (BBSRC)BB/S003835/1III - Bacteriology
173671Prof. R. Fitzgerald. Wellcome Trust Award 201531/Z/16/Z - Understanding bacterial host adaptation to combat infectious diseasesJose R PenadesWellcome Trust (WELLCOTR)201531/Z/16/ZInstitute of Infection, Immunity & Inflammation
172321DUT-SignalJose R PenadesEuropean Research Council (ERC)670932Institute of Infection, Immunity & Inflammation