Stage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening

dc.creatorKessler, Rafael Luis
dc.creatorGradia, Daniela Fiori
dc.creatorRampazzo, Rita de Cássia Pontello
dc.creatorLourenço, Edio Elıgio
dc.creatorFidêncio, Nilson José
dc.creatorManhaes, Lauro
dc.creatorProbst, Christian Macagnan
dc.creatorÁvila, Andréa Rodrigues
dc.creatorFragoso, Stenio Perdigão
dc.date2016-09-30T17:59:59Z
dc.date2016-09-30T17:59:59Z
dc.date2013
dc.date.accessioned2023-09-26T22:28:01Z
dc.date.available2023-09-26T22:28:01Z
dc.identifierKESSLER, 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.identifier1932-6203
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/16018
dc.identifier10.1371/journal.pone.0067441
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8879599
dc.descriptionCNPq, Fundação Araucaria, Fiocruz, Capes
dc.descriptionTrypanosoma 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.
dc.formatapplication/pdf
dc.languageeng
dc.publisherIratxe Puebla
dc.rightsopen access
dc.subjectTrypanosoma cruzi
dc.subjectTransgenic parasites
dc.subjectHost-parasite interaction
dc.subjectDrug development
dc.subjectInterações Hospedeiro-Parasita
dc.subjectAvaliação de Medicamentos
dc.titleStage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening
dc.typeArticle


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