A synthetic biology approach to assemble and reboot clinically relevant Pseudomonas aeruginosa tailed phages
ABSTRACTThe rise in the frequency of antibiotic resistance has made bacterial infections, specifically Pseudomonas aeruginosa, a cause for greater concern. Phage therapy is a promising solution that uses naturally isolated phages to treat bacterial infections. Ecological limitations, which stipulate...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2024-03-01
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| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.02897-23 |
| _version_ | 1797276721090658304 |
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| author | Thomas Ipoutcha Ratanachat Racharaks Stefanie Huttelmaier Cole J. Wilson Egon A. Ozer Erica M. Hartmann |
| author_facet | Thomas Ipoutcha Ratanachat Racharaks Stefanie Huttelmaier Cole J. Wilson Egon A. Ozer Erica M. Hartmann |
| author_sort | Thomas Ipoutcha |
| collection | DOAJ |
| description | ABSTRACTThe rise in the frequency of antibiotic resistance has made bacterial infections, specifically Pseudomonas aeruginosa, a cause for greater concern. Phage therapy is a promising solution that uses naturally isolated phages to treat bacterial infections. Ecological limitations, which stipulate a discrete host range and the inevitable evolution of resistance, may be overcome through a better understanding of phage biology and the utilization of engineered phages. In this study, we developed a synthetic biology approach to construct tailed phages that naturally target clinically relevant strains of Pseudomonas aeruginosa. As proof of concept, we successfully cloned and assembled the JG024 and DMS3 phage genomes in yeast using transformation-associated recombination cloning and rebooted these two phage genomes in two different strains of P. aeruginosa. We identified factors that affected phage reboot efficiency like the phage species or the presence of antiviral defense systems in the bacterial strain. We have successfully extended this method to two other phage species and observed that the method enables the reboot of phages that are naturally unable to infect the strain used for reboot. This research represents a critical step toward the construction of clinically relevant, engineered P. aeruginosa phages.IMPORTANCEPseudomonas aeruginosa is a bacterium responsible for severe infections and a common major complication in cystic fibrosis. The use of antibiotics to treat bacterial infections has become increasingly difficult as antibiotic resistance has become more prevalent. Phage therapy is an alternative solution that is already being used in some European countries, but its use is limited by the narrow host range due to the phage receptor specificity, the presence of antiviral defense systems in the bacterial strain, and the possible emergence of phage resistance. In this study, we demonstrate the use of a synthetic biology approach to construct and reboot clinically relevant P. aeruginosa tailed phages. This method enables a significant expansion of possibilities through the construction of engineered phages for therapy applications. |
| first_indexed | 2024-03-07T15:33:20Z |
| format | Article |
| id | doaj.art-1288574401724f25825fb69d28a8d1d3 |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-03-07T15:33:20Z |
| publishDate | 2024-03-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-1288574401724f25825fb69d28a8d1d32024-03-05T14:04:36ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972024-03-0112310.1128/spectrum.02897-23A synthetic biology approach to assemble and reboot clinically relevant Pseudomonas aeruginosa tailed phagesThomas Ipoutcha0Ratanachat Racharaks1Stefanie Huttelmaier2Cole J. Wilson3Egon A. Ozer4Erica M. Hartmann5Department of Civil and Environmental Engineering, Northwestern University, Evanston, Illinois, USADepartment of Civil and Environmental Engineering, Northwestern University, Evanston, Illinois, USADepartment of Civil and Environmental Engineering, Northwestern University, Evanston, Illinois, USADepartment of Civil and Environmental Engineering, Northwestern University, Evanston, Illinois, USADivision of Infectious Diseases, Department of Medicine, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Civil and Environmental Engineering, Northwestern University, Evanston, Illinois, USAABSTRACTThe rise in the frequency of antibiotic resistance has made bacterial infections, specifically Pseudomonas aeruginosa, a cause for greater concern. Phage therapy is a promising solution that uses naturally isolated phages to treat bacterial infections. Ecological limitations, which stipulate a discrete host range and the inevitable evolution of resistance, may be overcome through a better understanding of phage biology and the utilization of engineered phages. In this study, we developed a synthetic biology approach to construct tailed phages that naturally target clinically relevant strains of Pseudomonas aeruginosa. As proof of concept, we successfully cloned and assembled the JG024 and DMS3 phage genomes in yeast using transformation-associated recombination cloning and rebooted these two phage genomes in two different strains of P. aeruginosa. We identified factors that affected phage reboot efficiency like the phage species or the presence of antiviral defense systems in the bacterial strain. We have successfully extended this method to two other phage species and observed that the method enables the reboot of phages that are naturally unable to infect the strain used for reboot. This research represents a critical step toward the construction of clinically relevant, engineered P. aeruginosa phages.IMPORTANCEPseudomonas aeruginosa is a bacterium responsible for severe infections and a common major complication in cystic fibrosis. The use of antibiotics to treat bacterial infections has become increasingly difficult as antibiotic resistance has become more prevalent. Phage therapy is an alternative solution that is already being used in some European countries, but its use is limited by the narrow host range due to the phage receptor specificity, the presence of antiviral defense systems in the bacterial strain, and the possible emergence of phage resistance. In this study, we demonstrate the use of a synthetic biology approach to construct and reboot clinically relevant P. aeruginosa tailed phages. This method enables a significant expansion of possibilities through the construction of engineered phages for therapy applications.https://journals.asm.org/doi/10.1128/spectrum.02897-23phage therapysynthetic biologyPseudomonas aeruginosaphage reboot |
| spellingShingle | Thomas Ipoutcha Ratanachat Racharaks Stefanie Huttelmaier Cole J. Wilson Egon A. Ozer Erica M. Hartmann A synthetic biology approach to assemble and reboot clinically relevant Pseudomonas aeruginosa tailed phages Microbiology Spectrum phage therapy synthetic biology Pseudomonas aeruginosa phage reboot |
| title | A synthetic biology approach to assemble and reboot clinically relevant Pseudomonas aeruginosa tailed phages |
| title_full | A synthetic biology approach to assemble and reboot clinically relevant Pseudomonas aeruginosa tailed phages |
| title_fullStr | A synthetic biology approach to assemble and reboot clinically relevant Pseudomonas aeruginosa tailed phages |
| title_full_unstemmed | A synthetic biology approach to assemble and reboot clinically relevant Pseudomonas aeruginosa tailed phages |
| title_short | A synthetic biology approach to assemble and reboot clinically relevant Pseudomonas aeruginosa tailed phages |
| title_sort | synthetic biology approach to assemble and reboot clinically relevant pseudomonas aeruginosa tailed phages |
| topic | phage therapy synthetic biology Pseudomonas aeruginosa phage reboot |
| url | https://journals.asm.org/doi/10.1128/spectrum.02897-23 |
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