Attenuation of a pathogenic Mycoplasma strain by modification of the obg gene by using synthetic biology approaches
Mycoplasma species are responsible for several economically significant livestock diseases for which there is a need for new and improved vaccines. Most of the existing mycoplasma vaccines are attenuated strains that have been empirically obtained by serial passages or by chemical mutagenesis. The r...
| Autores principales: | , , , , , , , , , , , |
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| Formato: | Journal Article |
| Lenguaje: | Inglés |
| Publicado: |
American Society for Microbiology
2019
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| Materias: | |
| Acceso en línea: | https://hdl.handle.net/10568/106289 |
| _version_ | 1855520764137046016 |
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| author | Lartigue, C. Valverde Timana, Y. Labroussaa, F. Schieck, Elise G. Liljander, Anne M. Sacchini, Flavio Posthaus, H. Batailler, B. Sirand-Pugnet, P. Vashee, S. Jores, Joerg Blanchard, Alain |
| author_browse | Batailler, B. Blanchard, Alain Jores, Joerg Labroussaa, F. Lartigue, C. Liljander, Anne M. Posthaus, H. Sacchini, Flavio Schieck, Elise G. Sirand-Pugnet, P. Valverde Timana, Y. Vashee, S. |
| author_facet | Lartigue, C. Valverde Timana, Y. Labroussaa, F. Schieck, Elise G. Liljander, Anne M. Sacchini, Flavio Posthaus, H. Batailler, B. Sirand-Pugnet, P. Vashee, S. Jores, Joerg Blanchard, Alain |
| author_sort | Lartigue, C. |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | Mycoplasma species are responsible for several economically significant livestock diseases for which there is a need for new and improved vaccines. Most of the existing mycoplasma vaccines are attenuated strains that have been empirically obtained by serial passages or by chemical mutagenesis. The recent development of synthetic biology approaches has opened the way for the engineering of live mycoplasma vaccines. Using these tools, the essential GTPase-encoding gene obg was modified directly on the Mycoplasma mycoides subsp. capri genome cloned in yeast, reproducing mutations suspected to induce a temperature-sensitive (TS+) phenotype. After transplantation of modified genomes into a recipient cell, the phenotype of the resulting M. mycoides subsp. capri mutants was characterized. Single-point obg mutations did not result in a strong TS+ phenotype in M. mycoides subsp. capri, but a clone presenting three obg mutations was shown to grow with difficulty at temperatures of ≥40°C. This particular mutant was then tested in a caprine septicemia model of M. mycoides subsp. capri infection. Five out of eight goats infected with the parental strain had to be euthanized, in contrast to one out of eight goats infected with the obg mutant, demonstrating an attenuation of virulence in the mutant. Moreover, the strain isolated from the euthanized animal in the group infected with the obg mutant was shown to carry a reversion in the obg gene associated with the loss of the TS+ phenotype. This study demonstrates the feasibility of building attenuated strains of mycoplasma that could contribute to the design of novel vaccines with improved safety. IMPORTANCE Animal diseases due to mycoplasmas are a major cause of morbidity and mortality associated with economic losses for farmers all over the world. Currently used mycoplasma vaccines exhibit several drawbacks, including low efficacy, short time of protection, adverse reactions, and difficulty in differentiating infected from vaccinated animals. Therefore, there is a need for improved vaccines to control animal mycoplasmoses. Here, we used genome engineering tools derived from synthetic biology approaches to produce targeted mutations in the essential GTPase-encoding obg gene of Mycoplasma mycoides subsp. capri. Some of the resulting mutants exhibited a marked temperature-sensitive phenotype. The virulence of one of the obg mutants was evaluated in a caprine septicemia model and found to be strongly reduced. Although the obg mutant reverted to a virulent phenotype in one infected animal, we believe that these results contribute to a strategy that should help in building new vaccines against animal mycoplasmoses. |
| format | Journal Article |
| id | CGSpace106289 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2019 |
| publishDateRange | 2019 |
| publishDateSort | 2019 |
| publisher | American Society for Microbiology |
| publisherStr | American Society for Microbiology |
| record_format | dspace |
| spelling | CGSpace1062892025-12-08T09:54:28Z Attenuation of a pathogenic Mycoplasma strain by modification of the obg gene by using synthetic biology approaches Lartigue, C. Valverde Timana, Y. Labroussaa, F. Schieck, Elise G. Liljander, Anne M. Sacchini, Flavio Posthaus, H. Batailler, B. Sirand-Pugnet, P. Vashee, S. Jores, Joerg Blanchard, Alain mycoplasma genomes vaccines research animal diseases pathogenicity Mycoplasma species are responsible for several economically significant livestock diseases for which there is a need for new and improved vaccines. Most of the existing mycoplasma vaccines are attenuated strains that have been empirically obtained by serial passages or by chemical mutagenesis. The recent development of synthetic biology approaches has opened the way for the engineering of live mycoplasma vaccines. Using these tools, the essential GTPase-encoding gene obg was modified directly on the Mycoplasma mycoides subsp. capri genome cloned in yeast, reproducing mutations suspected to induce a temperature-sensitive (TS+) phenotype. After transplantation of modified genomes into a recipient cell, the phenotype of the resulting M. mycoides subsp. capri mutants was characterized. Single-point obg mutations did not result in a strong TS+ phenotype in M. mycoides subsp. capri, but a clone presenting three obg mutations was shown to grow with difficulty at temperatures of ≥40°C. This particular mutant was then tested in a caprine septicemia model of M. mycoides subsp. capri infection. Five out of eight goats infected with the parental strain had to be euthanized, in contrast to one out of eight goats infected with the obg mutant, demonstrating an attenuation of virulence in the mutant. Moreover, the strain isolated from the euthanized animal in the group infected with the obg mutant was shown to carry a reversion in the obg gene associated with the loss of the TS+ phenotype. This study demonstrates the feasibility of building attenuated strains of mycoplasma that could contribute to the design of novel vaccines with improved safety. IMPORTANCE Animal diseases due to mycoplasmas are a major cause of morbidity and mortality associated with economic losses for farmers all over the world. Currently used mycoplasma vaccines exhibit several drawbacks, including low efficacy, short time of protection, adverse reactions, and difficulty in differentiating infected from vaccinated animals. Therefore, there is a need for improved vaccines to control animal mycoplasmoses. Here, we used genome engineering tools derived from synthetic biology approaches to produce targeted mutations in the essential GTPase-encoding obg gene of Mycoplasma mycoides subsp. capri. Some of the resulting mutants exhibited a marked temperature-sensitive phenotype. The virulence of one of the obg mutants was evaluated in a caprine septicemia model and found to be strongly reduced. Although the obg mutant reverted to a virulent phenotype in one infected animal, we believe that these results contribute to a strategy that should help in building new vaccines against animal mycoplasmoses. 2019-06-26 2019-12-20T12:03:24Z 2019-12-20T12:03:24Z Journal Article https://hdl.handle.net/10568/106289 en Open Access American Society for Microbiology Lartigue, C., Valverde Timana, Y., Labroussaa, F., Schieck, E., Liljander, A., Sacchini, F., Posthaus, H., Batailler, B., Sirand-Pugnet, P., Vashee, S., Jores, J. and Blanchard, A. 2019. Attenuation of a pathogenic Mycoplasma strain by modification of the obg gene by using synthetic biology approaches. mSphere 4(3): e00030-19. |
| spellingShingle | mycoplasma genomes vaccines research animal diseases pathogenicity Lartigue, C. Valverde Timana, Y. Labroussaa, F. Schieck, Elise G. Liljander, Anne M. Sacchini, Flavio Posthaus, H. Batailler, B. Sirand-Pugnet, P. Vashee, S. Jores, Joerg Blanchard, Alain Attenuation of a pathogenic Mycoplasma strain by modification of the obg gene by using synthetic biology approaches |
| title | Attenuation of a pathogenic Mycoplasma strain by modification of the obg gene by using synthetic biology approaches |
| title_full | Attenuation of a pathogenic Mycoplasma strain by modification of the obg gene by using synthetic biology approaches |
| title_fullStr | Attenuation of a pathogenic Mycoplasma strain by modification of the obg gene by using synthetic biology approaches |
| title_full_unstemmed | Attenuation of a pathogenic Mycoplasma strain by modification of the obg gene by using synthetic biology approaches |
| title_short | Attenuation of a pathogenic Mycoplasma strain by modification of the obg gene by using synthetic biology approaches |
| title_sort | attenuation of a pathogenic mycoplasma strain by modification of the obg gene by using synthetic biology approaches |
| topic | mycoplasma genomes vaccines research animal diseases pathogenicity |
| url | https://hdl.handle.net/10568/106289 |
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