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

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 parasi...

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Autores principales:	Rafael Luis Kessler, Daniela Fiori Gradia, Rita de Cássia Pontello Rampazzo, Édio Elígio Lourenço, Nilson José Fidêncio, Lauro Manhaes, Christian Macagnan Probst, Andréa Rodrigues Ávila, Stenio Perdigão Fragoso
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/cb46d9c4c69d4cda9d3683e57d4b3cdc
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spelling	oai:doaj.org-article:cb46d9c4c69d4cda9d3683e57d4b3cdc2021-11-18T07:40:45ZStage-regulated GFP Expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening.1932-620310.1371/journal.pone.0067441https://doaj.org/article/cb46d9c4c69d4cda9d3683e57d4b3cdc2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23840703/?tool=EBIhttps://doaj.org/toc/1932-6203Trypanosoma 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.Rafael Luis KesslerDaniela Fiori GradiaRita de Cássia Pontello RampazzoÉdio Elígio LourençoNilson José FidêncioLauro ManhaesChristian Macagnan ProbstAndréa Rodrigues ÁvilaStenio Perdigão FragosoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 6, p e67441 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Rafael Luis Kessler Daniela Fiori Gradia Rita de Cássia Pontello Rampazzo Édio Elígio Lourenço Nilson José Fidêncio Lauro Manhaes Christian Macagnan Probst Andréa Rodrigues Ávila Stenio Perdigão Fragoso Stage-regulated GFP Expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening.
description	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.
format	article
author	Rafael Luis Kessler Daniela Fiori Gradia Rita de Cássia Pontello Rampazzo Édio Elígio Lourenço Nilson José Fidêncio Lauro Manhaes Christian Macagnan Probst Andréa Rodrigues Ávila Stenio Perdigão Fragoso
author_facet	Rafael Luis Kessler Daniela Fiori Gradia Rita de Cássia Pontello Rampazzo Édio Elígio Lourenço Nilson José Fidêncio Lauro Manhaes Christian Macagnan Probst Andréa Rodrigues Ávila Stenio Perdigão Fragoso
author_sort	Rafael Luis Kessler
title	Stage-regulated GFP Expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening.
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.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/cb46d9c4c69d4cda9d3683e57d4b3cdc
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Stage-regulated GFP Expression in Trypanosoma cruzi: applications from host-parasite interactions to drug screening.

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