Stage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2013 |
| Outros Autores: | , , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Institucional da FIOCRUZ (ARCA) |
| Texto Completo: | https://www.arca.fiocruz.br/handle/icict/16018 |
Resumo: | CNPq, Fundação Araucaria, Fiocruz, Capes |
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Kessler, Rafael LuisGradia, Daniela FioriRampazzo, Rita de Cássia PontelloLourenço, Edio ElıgioFidêncio, Nilson JoséManhaes, LauroProbst, Christian MacagnanÁvila, Andréa RodriguesFragoso, Stenio Perdigão2016-09-30T17:59:59Z2016-09-30T17:59:59Z2013KESSLER, Rafael Luis et al. Stage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening. PLoS ONE, v. 8, n. 6, p. 1-12, 2013.1932-6203https://www.arca.fiocruz.br/handle/icict/1601810.1371/journal.pone.0067441CNPq, Fundação Araucaria, Fiocruz, CapesFundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Trypanosoma cruzi is the etiological agent of Chagas disease, an illness that affects about 10 million people, mostly in South America, for which there is no effective treatment or vaccine. In this context, transgenic parasites expressing reporter genes are interesting tools for investigating parasite biology and host-parasite interactions, with a view to developing new strategies for disease prevention and treatment. We describe here the construction of a stably transfected fluorescent T. cruzi clone in which the GFP gene is integrated into the chromosome carrying the ribosomal cistron in T. cruzi Dm28c. This fluorescent T. cruzi produces detectable amounts of GFP only at replicative stages (epimastigote and amastigote), consistent with the larger amounts of GFP mRNA detected in these forms than in the non replicative trypomastigote stages. The fluorescence signal was also strongly correlated with the total number of parasites in T. cruzi cultures, providing a simple and rapid means of determining the growth inhibitory dose of anti-T.cruzi drugs in epimastigotes, by fluorometric microplate screening, and in amastigotes, by the flow cytometric quantification of T. cruzi-infected Vero cells. This fluorescent T. cruzi clone is, thus, an interesting tool for unbiased detection of the proliferating stages of the parasite, with multiple applications in the genetic analysis of T. cruzi, including analyses of host-parasite interactions, gene expression regulation and drug development.engIratxe PueblaStage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screeninginfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleTrypanosoma cruziTransgenic parasitesHost-parasite interactionDrug developmentInterações Hospedeiro-ParasitaAvaliação de Medicamentosinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-83074https://www.arca.fiocruz.br/bitstream/icict/16018/1/license.txt0271fb68963c017aa933c7aff60379f4MD51ORIGINALStage-Regulated GFP Expression in Trypanosoma cruzi.PDFStage-Regulated GFP Expression in Trypanosoma cruzi.PDFapplication/pdf2194571https://www.arca.fiocruz.br/bitstream/icict/16018/2/Stage-Regulated%20GFP%20Expression%20in%20Trypanosoma%20cruzi.PDF7ad80c423fdb2e2b11b74f6ec9b0b6b5MD52TEXTStage-Regulated GFP Expression in Trypanosoma cruzi.PDF.txtStage-Regulated GFP Expression in Trypanosoma cruzi.PDF.txtExtracted texttext/plain64651https://www.arca.fiocruz.br/bitstream/icict/16018/3/Stage-Regulated%20GFP%20Expression%20in%20Trypanosoma%20cruzi.PDF.txtd2aff59a8b7a44b4da9472a19b4e4862MD53icict/160182019-11-23 21:17:05.32oai:www.arca.fiocruz.br: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Repositório 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| dc.title.pt_BR.fl_str_mv |
Stage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening |
| title |
Stage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening |
| spellingShingle |
Stage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening Kessler, Rafael Luis Trypanosoma cruzi Transgenic parasites Host-parasite interaction Drug development Interações Hospedeiro-Parasita Avaliação de Medicamentos |
| title_short |
Stage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening |
| title_full |
Stage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening |
| title_fullStr |
Stage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening |
| title_full_unstemmed |
Stage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening |
| title_sort |
Stage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening |
| author |
Kessler, Rafael Luis |
| author_facet |
Kessler, Rafael Luis Gradia, Daniela Fiori Rampazzo, Rita de Cássia Pontello Lourenço, Edio Elıgio Fidêncio, Nilson José Manhaes, Lauro Probst, Christian Macagnan Ávila, Andréa Rodrigues Fragoso, Stenio Perdigão |
| author_role |
author |
| author2 |
Gradia, Daniela Fiori Rampazzo, Rita de Cássia Pontello Lourenço, Edio Elıgio Fidêncio, Nilson José Manhaes, Lauro Probst, Christian Macagnan Ávila, Andréa Rodrigues Fragoso, Stenio Perdigão |
| author2_role |
author author author author author author author author |
| dc.contributor.author.fl_str_mv |
Kessler, Rafael Luis Gradia, Daniela Fiori Rampazzo, Rita de Cássia Pontello Lourenço, Edio Elıgio Fidêncio, Nilson José Manhaes, Lauro Probst, Christian Macagnan Ávila, Andréa Rodrigues Fragoso, Stenio Perdigão |
| dc.subject.en.pt_BR.fl_str_mv |
Trypanosoma cruzi Transgenic parasites Host-parasite interaction Drug development |
| topic |
Trypanosoma cruzi Transgenic parasites Host-parasite interaction Drug development Interações Hospedeiro-Parasita Avaliação de Medicamentos |
| dc.subject.decs.pt_BR.fl_str_mv |
Interações Hospedeiro-Parasita Avaliação de Medicamentos |
| description |
CNPq, Fundação Araucaria, Fiocruz, Capes |
| publishDate |
2013 |
| dc.date.issued.fl_str_mv |
2013 |
| dc.date.accessioned.fl_str_mv |
2016-09-30T17:59:59Z |
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2016-09-30T17:59:59Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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publishedVersion |
| dc.identifier.citation.fl_str_mv |
KESSLER, Rafael Luis et al. Stage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening. PLoS ONE, v. 8, n. 6, p. 1-12, 2013. |
| dc.identifier.uri.fl_str_mv |
https://www.arca.fiocruz.br/handle/icict/16018 |
| dc.identifier.issn.pt_BR.fl_str_mv |
1932-6203 |
| dc.identifier.doi.pt_BR.fl_str_mv |
10.1371/journal.pone.0067441 |
| identifier_str_mv |
KESSLER, Rafael Luis et al. Stage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening. PLoS ONE, v. 8, n. 6, p. 1-12, 2013. 1932-6203 10.1371/journal.pone.0067441 |
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https://www.arca.fiocruz.br/handle/icict/16018 |
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Iratxe Puebla |
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Iratxe Puebla |
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