Removal of a subset of non-essential genes fully attenuates a highly virulent Mycoplasma strain
Mycoplasmas are the smallest free-living organisms and cause a number of economically important diseases affecting humans, animals, insects, and plants. Here, we demonstrate that highly virulent Mycoplasma mycoides subspecies capri (Mmc) can be fully attenuated via targeted deletion of non-essential...
| Autores principales: | , , , , , , , , , , , , , , , , |
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| Formato: | Journal Article |
| Lenguaje: | Inglés |
| Publicado: |
Frontiers Media
2019
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| Materias: | |
| Acceso en línea: | https://hdl.handle.net/10568/106284 |
| _version_ | 1855532354896920576 |
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| author | Jores, Joerg Ma, L. Ssajjakambwe, Paul Schieck, Elise G. Liljander, Anne M. Chandran, S. Stoffel, M.H. Cippa, V. Arfi, Y. Assad-Garcia, N. Falquet, L. Sirand-Pugnet, P. Blanchard, Alain Lartigue, C. Posthaus, H. Labroussaa, F. Vashee, S. |
| author_browse | Arfi, Y. Assad-Garcia, N. Blanchard, Alain Chandran, S. Cippa, V. Falquet, L. Jores, Joerg Labroussaa, F. Lartigue, C. Liljander, Anne M. Ma, L. Posthaus, H. Schieck, Elise G. Sirand-Pugnet, P. Ssajjakambwe, Paul Stoffel, M.H. Vashee, S. |
| author_facet | Jores, Joerg Ma, L. Ssajjakambwe, Paul Schieck, Elise G. Liljander, Anne M. Chandran, S. Stoffel, M.H. Cippa, V. Arfi, Y. Assad-Garcia, N. Falquet, L. Sirand-Pugnet, P. Blanchard, Alain Lartigue, C. Posthaus, H. Labroussaa, F. Vashee, S. |
| author_sort | Jores, Joerg |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | Mycoplasmas are the smallest free-living organisms and cause a number of economically important diseases affecting humans, animals, insects, and plants. Here, we demonstrate that highly virulent Mycoplasma mycoides subspecies capri (Mmc) can be fully attenuated via targeted deletion of non-essential genes encoding, among others, potential virulence traits. Five genomic regions, representing approximately 10% of the original Mmc genome, were successively deleted using Saccharomyces cerevisiae as an engineering platform. Specifically, a total of 68 genes out of the 432 genes verified to be individually non-essential in the JCVI-Syn3.0 minimal cell, were excised from the genome. In vitro characterization showed that this mutant was similar to its parental strain in terms of its doubling time, even though 10% of the genome content were removed. A novel in vivo challenge model in goats revealed that the wild-type parental strain caused marked necrotizing inflammation at the site of inoculation, septicemia and all animals reached endpoint criteria within 6 days after experimental infection. This is in contrast to the mutant strain, which caused no clinical signs nor pathomorphological lesions. These results highlight, for the first time, the rational design, construction and complete attenuation of a Mycoplasma strain via synthetic genomics tools. Trait addition using the yeast-based genome engineering platform and subsequent in vitro or in vivo trials employing the Mycoplasma chassis will allow us to dissect the role of individual candidate Mycoplasma virulence factors and lead the way for the development of an attenuated designer vaccine. |
| format | Journal Article |
| id | CGSpace106284 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2019 |
| publishDateRange | 2019 |
| publishDateSort | 2019 |
| publisher | Frontiers Media |
| publisherStr | Frontiers Media |
| record_format | dspace |
| spelling | CGSpace1062842025-12-08T09:54:28Z Removal of a subset of non-essential genes fully attenuates a highly virulent Mycoplasma strain Jores, Joerg Ma, L. Ssajjakambwe, Paul Schieck, Elise G. Liljander, Anne M. Chandran, S. Stoffel, M.H. Cippa, V. Arfi, Y. Assad-Garcia, N. Falquet, L. Sirand-Pugnet, P. Blanchard, Alain Lartigue, C. Posthaus, H. Labroussaa, F. Vashee, S. mycoplasma genomes virulence research animal diseases Mycoplasmas are the smallest free-living organisms and cause a number of economically important diseases affecting humans, animals, insects, and plants. Here, we demonstrate that highly virulent Mycoplasma mycoides subspecies capri (Mmc) can be fully attenuated via targeted deletion of non-essential genes encoding, among others, potential virulence traits. Five genomic regions, representing approximately 10% of the original Mmc genome, were successively deleted using Saccharomyces cerevisiae as an engineering platform. Specifically, a total of 68 genes out of the 432 genes verified to be individually non-essential in the JCVI-Syn3.0 minimal cell, were excised from the genome. In vitro characterization showed that this mutant was similar to its parental strain in terms of its doubling time, even though 10% of the genome content were removed. A novel in vivo challenge model in goats revealed that the wild-type parental strain caused marked necrotizing inflammation at the site of inoculation, septicemia and all animals reached endpoint criteria within 6 days after experimental infection. This is in contrast to the mutant strain, which caused no clinical signs nor pathomorphological lesions. These results highlight, for the first time, the rational design, construction and complete attenuation of a Mycoplasma strain via synthetic genomics tools. Trait addition using the yeast-based genome engineering platform and subsequent in vitro or in vivo trials employing the Mycoplasma chassis will allow us to dissect the role of individual candidate Mycoplasma virulence factors and lead the way for the development of an attenuated designer vaccine. 2019-04-03 2019-12-20T11:28:54Z 2019-12-20T11:28:54Z Journal Article https://hdl.handle.net/10568/106284 en Open Access Frontiers Media Jores, J., Ma, L., Ssajjakambwe, P., Schieck, E., Liljander, A., Chandran, S., Stoffel, M.H., Cippa, V., Arfi, Y., Assad-Garcia, N., Falquet, L., Sirand-Pugnet, P., Blanchard, A., Lartigue, C., Posthaus, H., Labroussaa, F. and Vashee, S. 2019. Removal of a subset of non-essential genes fully attenuates a highly virulent Mycoplasma strain. Frontiers in Microbiology 10: 664. |
| spellingShingle | mycoplasma genomes virulence research animal diseases Jores, Joerg Ma, L. Ssajjakambwe, Paul Schieck, Elise G. Liljander, Anne M. Chandran, S. Stoffel, M.H. Cippa, V. Arfi, Y. Assad-Garcia, N. Falquet, L. Sirand-Pugnet, P. Blanchard, Alain Lartigue, C. Posthaus, H. Labroussaa, F. Vashee, S. Removal of a subset of non-essential genes fully attenuates a highly virulent Mycoplasma strain |
| title | Removal of a subset of non-essential genes fully attenuates a highly virulent Mycoplasma strain |
| title_full | Removal of a subset of non-essential genes fully attenuates a highly virulent Mycoplasma strain |
| title_fullStr | Removal of a subset of non-essential genes fully attenuates a highly virulent Mycoplasma strain |
| title_full_unstemmed | Removal of a subset of non-essential genes fully attenuates a highly virulent Mycoplasma strain |
| title_short | Removal of a subset of non-essential genes fully attenuates a highly virulent Mycoplasma strain |
| title_sort | removal of a subset of non essential genes fully attenuates a highly virulent mycoplasma strain |
| topic | mycoplasma genomes virulence research animal diseases |
| url | https://hdl.handle.net/10568/106284 |
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