Complete in vitro life cycle of Trypanosoma congolense: development of genetic tools.
BACKGROUND: Animal African trypanosomosis, a disease mainly caused by the protozoan parasite Trypanosoma congolense, is a major constraint to livestock productivity and has a significant impact in the developing countries of Africa. RNA interference (RNAi) has been used to study gene function and id...
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ftdoajarticles:oai:doaj.org/article:7d1fb77590574b69acd93715ea81f276 2023-05-15T15:12:03+02:00 Complete in vitro life cycle of Trypanosoma congolense: development of genetic tools. Virginie Coustou Fabien Guegan Nicolas Plazolles Théo Baltz 2010-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0000618 https://doaj.org/article/7d1fb77590574b69acd93715ea81f276 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC2830455?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0000618 https://doaj.org/article/7d1fb77590574b69acd93715ea81f276 PLoS Neglected Tropical Diseases, Vol 4, Iss 3, p e618 (2010) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2010 ftdoajarticles https://doi.org/10.1371/journal.pntd.0000618 2022-12-31T03:53:47Z BACKGROUND: Animal African trypanosomosis, a disease mainly caused by the protozoan parasite Trypanosoma congolense, is a major constraint to livestock productivity and has a significant impact in the developing countries of Africa. RNA interference (RNAi) has been used to study gene function and identify drug and vaccine targets in a variety of organisms including trypanosomes. However, trypanosome RNAi studies have mainly been conducted in T. brucei, as a model for human infection, largely ignoring livestock parasites of economical importance such as T. congolense, which displays different pathogenesis profiles. The whole T. congolense life cycle can be completed in vitro, but this attractive model displayed important limitations: (i) genetic tools were currently limited to insect forms and production of modified infectious BSF through differentiation was never achieved, (ii) in vitro differentiation techniques lasted several months, (iii) absence of long-term bloodstream forms (BSF) in vitro culture prevented genomic analyses. METHODOLOGY/PRINCIPAL FINDINGS: We optimized culture conditions for each developmental stage and secured the differentiation steps. Specifically, we devised a medium adapted for the strenuous development of stable long-term BSF culture. Using Amaxa nucleofection technology, we greatly improved the transfection rate of the insect form and designed an inducible transgene expression system using the IL3000 reference strain. We tested it by expression of reporter genes and through RNAi. Subsequently, we achieved the complete in vitro life cycle with dramatically shortened time requirements for various wild type and transgenic strains. Finally, we established the use of modified strains for experimental infections and underlined a host adaptation phase requirement. CONCLUSIONS/SIGNIFICANCE: We devised an improved T. congolense model, which offers the opportunity to perform functional genomics analyses throughout the whole life cycle. It represents a very useful tool to understand ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLoS Neglected Tropical Diseases 4 3 e618 |
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Open Polar |
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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 Virginie Coustou Fabien Guegan Nicolas Plazolles Théo Baltz Complete in vitro life cycle of Trypanosoma congolense: development of genetic tools. |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
BACKGROUND: Animal African trypanosomosis, a disease mainly caused by the protozoan parasite Trypanosoma congolense, is a major constraint to livestock productivity and has a significant impact in the developing countries of Africa. RNA interference (RNAi) has been used to study gene function and identify drug and vaccine targets in a variety of organisms including trypanosomes. However, trypanosome RNAi studies have mainly been conducted in T. brucei, as a model for human infection, largely ignoring livestock parasites of economical importance such as T. congolense, which displays different pathogenesis profiles. The whole T. congolense life cycle can be completed in vitro, but this attractive model displayed important limitations: (i) genetic tools were currently limited to insect forms and production of modified infectious BSF through differentiation was never achieved, (ii) in vitro differentiation techniques lasted several months, (iii) absence of long-term bloodstream forms (BSF) in vitro culture prevented genomic analyses. METHODOLOGY/PRINCIPAL FINDINGS: We optimized culture conditions for each developmental stage and secured the differentiation steps. Specifically, we devised a medium adapted for the strenuous development of stable long-term BSF culture. Using Amaxa nucleofection technology, we greatly improved the transfection rate of the insect form and designed an inducible transgene expression system using the IL3000 reference strain. We tested it by expression of reporter genes and through RNAi. Subsequently, we achieved the complete in vitro life cycle with dramatically shortened time requirements for various wild type and transgenic strains. Finally, we established the use of modified strains for experimental infections and underlined a host adaptation phase requirement. CONCLUSIONS/SIGNIFICANCE: We devised an improved T. congolense model, which offers the opportunity to perform functional genomics analyses throughout the whole life cycle. It represents a very useful tool to understand ... |
format |
Article in Journal/Newspaper |
author |
Virginie Coustou Fabien Guegan Nicolas Plazolles Théo Baltz |
author_facet |
Virginie Coustou Fabien Guegan Nicolas Plazolles Théo Baltz |
author_sort |
Virginie Coustou |
title |
Complete in vitro life cycle of Trypanosoma congolense: development of genetic tools. |
title_short |
Complete in vitro life cycle of Trypanosoma congolense: development of genetic tools. |
title_full |
Complete in vitro life cycle of Trypanosoma congolense: development of genetic tools. |
title_fullStr |
Complete in vitro life cycle of Trypanosoma congolense: development of genetic tools. |
title_full_unstemmed |
Complete in vitro life cycle of Trypanosoma congolense: development of genetic tools. |
title_sort |
complete in vitro life cycle of trypanosoma congolense: development of genetic tools. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2010 |
url |
https://doi.org/10.1371/journal.pntd.0000618 https://doaj.org/article/7d1fb77590574b69acd93715ea81f276 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 4, Iss 3, p e618 (2010) |
op_relation |
http://europepmc.org/articles/PMC2830455?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0000618 https://doaj.org/article/7d1fb77590574b69acd93715ea81f276 |
op_doi |
https://doi.org/10.1371/journal.pntd.0000618 |
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PLoS Neglected Tropical Diseases |
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4 |
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3 |
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e618 |
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