Rebooting Synthetic Phage-Inducible Chromosomal Islands: One Method to Forge Them All
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....
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
American Association for the Advancement of Science (AAAS)
2020-01-01
|
| Series: | BioDesign Research |
| Online Access: | http://dx.doi.org/10.34133/2020/5783064 |
| _version_ | 1827337525115486208 |
|---|---|
| author | Rodrigo Ibarra-Chávez Andreas F. Haag Pedro Dorado-Morales Iñigo Lasa José R. Penadés |
| author_facet | Rodrigo Ibarra-Chávez Andreas F. Haag Pedro Dorado-Morales Iñigo Lasa José R. Penadés |
| author_sort | Rodrigo Ibarra-Chávez |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-07T18:57:22Z |
| format | Article |
| id | doaj.art-684a0cf7f5194c44843d526ead44e035 |
| institution | Directory Open Access Journal |
| issn | 2693-1257 |
| language | English |
| last_indexed | 2024-03-07T18:57:22Z |
| publishDate | 2020-01-01 |
| publisher | American Association for the Advancement of Science (AAAS) |
| record_format | Article |
| series | BioDesign Research |
| spelling | doaj.art-684a0cf7f5194c44843d526ead44e0352024-03-02T00:08:56ZengAmerican Association for the Advancement of Science (AAAS)BioDesign Research2693-12572020-01-01202010.34133/2020/5783064Rebooting Synthetic Phage-Inducible Chromosomal Islands: One Method to Forge Them AllRodrigo Ibarra-Chávez0Andreas F. Haag1Pedro Dorado-Morales2Iñigo Lasa3José R. Penadés4Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UKInstitute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UKLaboratory of Microbial Pathogenesis, Navarrabiomed, Complejo Hospitalario de Navarra-Universidad Pública de Navarra (UPNA), Instituto de Investigación Sanitaria de Navarra (IDISNA), 31008 Pamplona, SpainLaboratory of Microbial Pathogenesis, Navarrabiomed, Complejo Hospitalario de Navarra-Universidad Pública de Navarra (UPNA), Instituto de Investigación Sanitaria de Navarra (IDISNA), 31008 Pamplona, SpainInstitute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UKPhage-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.http://dx.doi.org/10.34133/2020/5783064 |
| spellingShingle | Rodrigo Ibarra-Chávez Andreas F. Haag Pedro Dorado-Morales Iñigo Lasa José R. Penadés Rebooting Synthetic Phage-Inducible Chromosomal Islands: One Method to Forge Them All BioDesign Research |
| title | Rebooting Synthetic Phage-Inducible Chromosomal Islands: One Method to Forge Them All |
| title_full | Rebooting Synthetic Phage-Inducible Chromosomal Islands: One Method to Forge Them All |
| title_fullStr | Rebooting Synthetic Phage-Inducible Chromosomal Islands: One Method to Forge Them All |
| title_full_unstemmed | Rebooting Synthetic Phage-Inducible Chromosomal Islands: One Method to Forge Them All |
| title_short | Rebooting Synthetic Phage-Inducible Chromosomal Islands: One Method to Forge Them All |
| title_sort | rebooting synthetic phage inducible chromosomal islands one method to forge them all |
| url | http://dx.doi.org/10.34133/2020/5783064 |
| work_keys_str_mv | AT rodrigoibarrachx00e1vez rebootingsyntheticphageinduciblechromosomalislandsonemethodtoforgethemall AT andreasfhaag rebootingsyntheticphageinduciblechromosomalislandsonemethodtoforgethemall AT pedrodoradomorales rebootingsyntheticphageinduciblechromosomalislandsonemethodtoforgethemall AT ix00f1igolasa rebootingsyntheticphageinduciblechromosomalislandsonemethodtoforgethemall AT josx00e9rpenadx00e9s rebootingsyntheticphageinduciblechromosomalislandsonemethodtoforgethemall |