Development and validation of systems for genetic manipulation of the Old World tick-borne relapsing fever spirochete, Borrelia duttonii.
Relapsing fever (RF), a vector-borne disease caused by Borrelia spp., is characterized by recurring febrile episodes due to repeated bouts of bacteremia. RF spirochetes can be geographically and phylogenetically divided into two distinct groups; Old World RF Borrelia (found in Africa, Asia, and Euro...
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ftdoajarticles:oai:doaj.org/article:96b34f5dc8854deaa9812a5f61955f9a 2024-09-09T19:27:12+00:00 Development and validation of systems for genetic manipulation of the Old World tick-borne relapsing fever spirochete, Borrelia duttonii. Clay D Jackson-Litteken Wanfeng Guo Brandon A Hogland C Tyler Ratliff LeAnn McFadden Marissa S Fullerton Daniel E Voth Ryan O M Rego Jon S Blevins 2024-07-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0012348 https://doaj.org/article/96b34f5dc8854deaa9812a5f61955f9a EN eng Public Library of Science (PLoS) https://doi.org/10.1371/journal.pntd.0012348 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0012348 https://doaj.org/article/96b34f5dc8854deaa9812a5f61955f9a PLoS Neglected Tropical Diseases, Vol 18, Iss 7, p e0012348 (2024) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2024 ftdoajarticles https://doi.org/10.1371/journal.pntd.0012348 2024-08-19T14:56:40Z Relapsing fever (RF), a vector-borne disease caused by Borrelia spp., is characterized by recurring febrile episodes due to repeated bouts of bacteremia. RF spirochetes can be geographically and phylogenetically divided into two distinct groups; Old World RF Borrelia (found in Africa, Asia, and Europe) and New World RF Borrelia (found in the Americas). While RF is a rarely reported disease in the Americas, RF is prevalent in endemic parts of Africa. Despite phylogenetic differences between Old World and New World RF Borrelia and higher incidence of disease associated with Old World RF spirochete infection, genetic manipulation has only been described in New World RF bacteria. Herein, we report the generation of genetic tools for use in the Old World RF spirochete, Borrelia duttonii. We describe methods for transformation and establish shuttle vector- and integration-based approaches for genetic complementation, creating green fluorescent protein (gfp)-expressing B. duttonii strains as a proof of principle. Allelic exchange mutagenesis was also used to inactivate a homolog of the Borrelia burgdorferi p66 gene, which encodes an important virulence factor, in B. duttonii and demonstrate that this mutant was attenuated in a murine model of RF. Finally, the B. duttonii p66 mutant was complemented using shuttle vector- and cis integration-based approaches. As expected, complemented p66 mutant strains were fully infectious, confirming that P66 is required for optimal mammalian infection. The genetic tools and techniques reported herein represent an important advancement in the study of RF Borrelia that allows for future characterization of virulence determinants and colonization factors important for the enzootic cycle of Old World RF spirochetes. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 18 7 e0012348 |
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Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Clay D Jackson-Litteken Wanfeng Guo Brandon A Hogland C Tyler Ratliff LeAnn McFadden Marissa S Fullerton Daniel E Voth Ryan O M Rego Jon S Blevins Development and validation of systems for genetic manipulation of the Old World tick-borne relapsing fever spirochete, Borrelia duttonii. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Relapsing fever (RF), a vector-borne disease caused by Borrelia spp., is characterized by recurring febrile episodes due to repeated bouts of bacteremia. RF spirochetes can be geographically and phylogenetically divided into two distinct groups; Old World RF Borrelia (found in Africa, Asia, and Europe) and New World RF Borrelia (found in the Americas). While RF is a rarely reported disease in the Americas, RF is prevalent in endemic parts of Africa. Despite phylogenetic differences between Old World and New World RF Borrelia and higher incidence of disease associated with Old World RF spirochete infection, genetic manipulation has only been described in New World RF bacteria. Herein, we report the generation of genetic tools for use in the Old World RF spirochete, Borrelia duttonii. We describe methods for transformation and establish shuttle vector- and integration-based approaches for genetic complementation, creating green fluorescent protein (gfp)-expressing B. duttonii strains as a proof of principle. Allelic exchange mutagenesis was also used to inactivate a homolog of the Borrelia burgdorferi p66 gene, which encodes an important virulence factor, in B. duttonii and demonstrate that this mutant was attenuated in a murine model of RF. Finally, the B. duttonii p66 mutant was complemented using shuttle vector- and cis integration-based approaches. As expected, complemented p66 mutant strains were fully infectious, confirming that P66 is required for optimal mammalian infection. The genetic tools and techniques reported herein represent an important advancement in the study of RF Borrelia that allows for future characterization of virulence determinants and colonization factors important for the enzootic cycle of Old World RF spirochetes. |
format |
Article in Journal/Newspaper |
author |
Clay D Jackson-Litteken Wanfeng Guo Brandon A Hogland C Tyler Ratliff LeAnn McFadden Marissa S Fullerton Daniel E Voth Ryan O M Rego Jon S Blevins |
author_facet |
Clay D Jackson-Litteken Wanfeng Guo Brandon A Hogland C Tyler Ratliff LeAnn McFadden Marissa S Fullerton Daniel E Voth Ryan O M Rego Jon S Blevins |
author_sort |
Clay D Jackson-Litteken |
title |
Development and validation of systems for genetic manipulation of the Old World tick-borne relapsing fever spirochete, Borrelia duttonii. |
title_short |
Development and validation of systems for genetic manipulation of the Old World tick-borne relapsing fever spirochete, Borrelia duttonii. |
title_full |
Development and validation of systems for genetic manipulation of the Old World tick-borne relapsing fever spirochete, Borrelia duttonii. |
title_fullStr |
Development and validation of systems for genetic manipulation of the Old World tick-borne relapsing fever spirochete, Borrelia duttonii. |
title_full_unstemmed |
Development and validation of systems for genetic manipulation of the Old World tick-borne relapsing fever spirochete, Borrelia duttonii. |
title_sort |
development and validation of systems for genetic manipulation of the old world tick-borne relapsing fever spirochete, borrelia duttonii. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2024 |
url |
https://doi.org/10.1371/journal.pntd.0012348 https://doaj.org/article/96b34f5dc8854deaa9812a5f61955f9a |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 18, Iss 7, p e0012348 (2024) |
op_relation |
https://doi.org/10.1371/journal.pntd.0012348 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0012348 https://doaj.org/article/96b34f5dc8854deaa9812a5f61955f9a |
op_doi |
https://doi.org/10.1371/journal.pntd.0012348 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
18 |
container_issue |
7 |
container_start_page |
e0012348 |
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1809896664374181888 |