Author | Silva-Gomes, Natália Lins | |
Author | Rampazzo, Rita de Cássia Pontello | |
Author | Moreira, Claudia Maria do Nascimento | |
Author | Porcino, Gabriane Nascimento | |
Author | Santos, Cyndia Mara Bezerra dos | |
Author | Krieger, Marco Aurélio | |
Author | Vasconcelos, Eveline Gomes | |
Author | Fragoso, Stenio Perdigão | |
Author | Moreira, Otacilio C. | |
Access date | 2020-04-21T15:35:39Z | |
Available date | 2020-04-21T15:35:39Z | |
Document date | 2020 | |
Citation | SILVA-GOMES, Natália Lins et al. Knocking Down TcNTPDase-1 Gene Reduces in vitro Infectivity of Trypanosoma cruzi. Frontiers in Immunology, v. 11, n. 434, p. 1-12, 2020. | pt_BR |
ISSN | 1664-302X | pt_BR |
URI | https://www.arca.fiocruz.br/handle/icict/40905 | |
Language | por | pt_BR |
Rights | open access | pt_BR |
Subject in Portuguese | TcNTPDase-1 | pt_BR |
Title | Knocking down TcNTPDase-1 gene reduces in vitro infectivity of Trypanosoma cruzi | pt_BR |
Type | Article | pt_BR |
DOI | 10.3389/fmicb.2020.00434 | |
Abstract | Ecto-Nucleoside Triphosphate Diphosphohydrolases are enzymes that hydrolyze tri- and/or diphosphate nucleosides. Evidences pointed out to their participation in Trypanosoma cruzi virulence, infectivity, and purine acquisition. In this study, recombinant T. cruzi knocking out or overexpressing the TcNTPDase-1 gene were built, and the role of TcNTPDase-1 in the in vitro interaction with VERO cells was investigated. Results show that epimastigote forms of hemi-knockout parasites showed about 50% lower level of TcNTPDase-1 gene expression when compared to the wild type, while the T. cruzi overexpressing this gene reach 20 times higher gene expression. In trypomastigote forms, the same decreasing in TcNTPDase-1 gene expression was observed to the hemi-knockout parasites. The in vitro infection assays showed a reduction to 51.6 and 59.9% at the adhesion and to 25.2 and 26.4% at the endocytic indexes to the parasites knockout to one or other allele (Hygro and Neo hemi-knockouts), respectively. In contrast, the infection assays with T. cruzi overexpressing TcNTPDase-1 from the WT or Neo hemi-knockout parasites showed an opposite result, with the increasing to 287.7 and 271.1% at the adhesion and to 220.4 and 186.7% at the endocytic indexes, respectively. The parasitic load estimated in infected VERO cells by quantitative real time PCR corroborated these findings. Taken together, the partial silencing and overexpression of the TcNTPDase-1 gene generated viable parasites with low and high infectivity rates, respectively, corroborating that the enzyme encoded for this gene plays an important role to the T. cruzi infectivity. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Genômica Funcional. Curitiba, PR, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Molecular de Tripanossomatídeos. Curitiba, PR, Brasil. | pt_BR |
Affilliation | Universidade Federal de Juiz de Fora. Instituto de Ciências Biológicas. Laboratório de Estrutura e Função de Proteínas. Juiz de Fora, MG, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Genômica Funcional. Curitiba, PR, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Genômica Funcional. Curitiba, PR, Brasil. | pt_BR |
Affilliation | Universidade Federal de Juiz de Fora. Instituto de Ciências Biológicas. Laboratório de Estrutura e Função de Proteínas. Juiz de Fora, MG, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Molecular de Tripanossomatídeos. Curitiba, PR, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil. | pt_BR |
Subject | Gene Knockout Techniques | pt_BR |
Subject | pathogenicity | pt_BR |
Subject | infectivity | pt_BR |
Subject | Enzyme Activation | pt_BR |
Subject in Spanish | Técnicas de Inactivación de Genes | pt_BR |
Subject in Spanish | patogenicidad | pt_BR |
Subject in Spanish | Activación Enzimática | pt_BR |
DeCS | Trypanosoma cruzi | pt_BR |
DeCS | Técnicas de Inativação de Genes | pt_BR |
DeCS | patogenicidade | pt_BR |
DeCS | Ativação Enzimática | pt_BR |