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Removal of a subset of non-essential genes fully attenuates a highly virulent mycoplasma strain

  • Jores, Joerg Department of Infectious Diseases and Pathobiology, Institute of Veterinary Bacteriology, University of Bern, Switzerland - International Livestock Research Institute, Nairobi, Kenya
  • Ma, Li J. Craig Venter Institute, Rockville, MD, United States
  • Ssajjakambwe, Paul International Livestock Research Institute, Nairobi, Kenya - College of Veterinary Medicine, Animal Resources and Biosecurity, Makerere University, Kampala, Uganda
  • Schieck, Elise International Livestock Research Institute, Nairobi, Kenya
  • Liljander, Anne International Livestock Research Institute, Nairobi, Kenya
  • Chandran, Suchismita J. Craig Venter Institute, Rockville, MD, United States
  • Stoffel, Michael H. Division of Veterinary Anatomy, Department of Clinical Research and Veterinary Public Health, University of Bern, Switzerland
  • Cippa, Valentina Department of Infectious Diseases and Pathobiology, Institute of Veterinary Bacteriology, University of Bern, Switzerland
  • Arfi, Yonathan UMR 1332 – Biologie du Fruit et Pathologie, Institut National de la Recherche Agronomique, Villenave-d’Ornon, France - UMR 1332 – Biologie du Fruit et Pathologie, Université de Bordeaux, Villenave-d’Ornon, France
  • Assad-Garcia, Nacyra J. Craig Venter Institute, Rockville, MD, United States
  • Falquet, Laurent Biochemistry Unit, Swiss Institute of Bioinformatics, University of Fribourg, Switzerland
  • Sirand-Pugnet, Pascal UMR 1332 – Biologie du Fruit et Pathologie, Institut National de la Recherche Agronomique, Villenave-d’Ornon, France - UMR 1332 – Biologie du Fruit et Pathologie, Université de Bordeaux, Villenave-d’Ornon, France
  • Blanchard, Alain UMR 1332 – Biologie du Fruit et Pathologie, Institut National de la Recherche Agronomique, Villenave-d’Ornon, France - UMR 1332 – Biologie du Fruit et Pathologie, Université de Bordeaux, Villenave-d’Ornon, France
  • Lartigue, Carole UMR 1332 – Biologie du Fruit et Pathologie, Institut National de la Recherche Agronomique, Villenave-d’Ornon, France - UMR 1332 – Biologie du Fruit et Pathologie, Université de Bordeaux, Villenave-d’Ornon, France
  • Posthaus, Horst Department for Infectious Diseases and Pathobiology, Institute of Animal Pathology (COMPATH), University of Bern, Switzerland
  • Labroussaa, Fabien Department of Infectious Diseases and Pathobiology, Institute of Veterinary Bacteriology, University of Bern, Switzerland
  • Vashee, Sanjay J. Craig Venter Institute, Rockville, MD, United States
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  • 03.04.2019
Published in:
  • Frontiers in Microbiology. - 2019, vol. 10, p. 664
English 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.
Faculty
Faculté des sciences et de médecine
Department
Département de Biologie
Language
  • English
Classification
Biological sciences
License
License undefined
Identifiers
Persistent URL
https://folia.unifr.ch/unifr/documents/307819
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