Removal of a Subset of Non-essential Genes Fully Attenuates a Highly Virulent Mycoplasma Strain.

Jores, Jörg; Ma, Li; Ssajjakambwe, Paul; Schieck, Elise; Liljander, Anne; Chandran, Suchismita; Stoffel, Michael H.; Cippà, Valentina; Arfi, Yonathan; Assad-Garcia, Nacyra; Falquet, Laurent; Sirand-Pugnet, Pascal; Blanchard, Alain; Lartigue, Carole; Posthaus, Horst; Labroussaa, Fabien; Vashee, Sanjay (2019). Removal of a Subset of Non-essential Genes Fully Attenuates a Highly Virulent Mycoplasma Strain. Frontiers in Microbiology, 10(664), p. 664. Frontiers 10.3389/fmicb.2019.00664

[img]
Preview
Text
b140649.pdf - Published Version
Available under License Creative Commons: Attribution (CC-BY).

Download (8MB) | Preview

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.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Research Foci > Host-Pathogen Interaction
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Animal Pathology
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH) > Veterinary Anatomy
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Veterinary Bacteriology

UniBE Contributor:

Jores, Jörg; Stoffel, Michael Hubert; Cippà, Valentina Lucia; Posthaus, Horst and Labroussaa, Fabien

Subjects:

500 Science > 570 Life sciences; biology
600 Technology > 610 Medicine & health
600 Technology > 630 Agriculture

ISSN:

1664-302X

Publisher:

Frontiers

Language:

English

Submitter:

Achim Braun Parham

Date Deposited:

11 Mar 2020 12:56

Last Modified:

15 Mar 2020 02:49

Publisher DOI:

10.3389/fmicb.2019.00664

PubMed ID:

31001234

Uncontrolled Keywords:

Mycoplasma mycoides subsp. capri attenuation genome engineering in vivo challenge virulence traits

BORIS DOI:

10.7892/boris.140649

URI:

https://boris.unibe.ch/id/eprint/140649

Actions (login required)

Edit item Edit item
Provide Feedback