Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/16018
Title: Stage-regulated GFP expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening
Authors: Kessler, Rafael Luis
Gradia, Daniela Fiori
Rampazzo, Rita de Cassia Pontello
Lourenço, Edio Elıgio
Fidêncio, Nilson José
Manhaes, Lauro
Probst, Christian Macagnan
Ávila, Andréa Rodrigues
Fragoso, Stenio Perdigão
Affilliation: 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.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Abstract: 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.
Keywords: Trypanosoma cruzi
transgenic parasites
host-parasite interaction
drug development
DeCS: Interações Hospedeiro-Parasita
Avaliação de Medicamentos
Issue Date: 2013
Publisher: Iratxe Puebla
Citation: 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.
DOI: 10.1371/journal.pone.0067441
ISSN: 1932-6203
Copyright: open access
Appears in Collections:PR - ICC - Artigos de Periódicos

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