Knocking down TcNTPDase-1 gene reduces in vitro infectivity of Trypanosoma cruzi
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2020 |
| Outros Autores: | , , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | por |
| Título da fonte: | Repositório Institucional da FIOCRUZ (ARCA) |
| Texto Completo: | https://www.arca.fiocruz.br/handle/icict/40905 |
Resumo: | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil. |
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Silva-Gomes, Natália LinsRampazzo, Rita de Cássia PontelloMoreira, Claudia Maria do NascimentoPorcino, Gabriane NascimentoSantos, Cyndia Mara Bezerra dosKrieger, Marco AurélioVasconcelos, Eveline GomesFragoso, Stenio PerdigãoMoreira, Otacilio C.2020-04-21T15:35:39Z2020-04-21T15:35:39Z2020SILVA-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, 2020.1664-302Xhttps://www.arca.fiocruz.br/handle/icict/4090510.3389/fmicb.2020.00434porTcNTPDase-1Gene Knockout TechniquespathogenicityinfectivityEnzyme ActivationTécnicas de Inactivación de GenespatogenicidadActivación EnzimáticaTrypanosoma cruziTécnicas de Inativação de GenespatogenicidadeAtivação EnzimáticaKnocking down TcNTPDase-1 gene reduces in vitro infectivity of Trypanosoma cruziinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Genômica Funcional. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Molecular de Tripanossomatídeos. Curitiba, PR, Brasil.Universidade Federal de Juiz de Fora. Instituto de Ciências Biológicas. Laboratório de Estrutura e Função de Proteínas. Juiz de Fora, MG, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Genômica Funcional. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Genômica Funcional. Curitiba, PR, Brasil.Universidade Federal de Juiz de Fora. Instituto de Ciências Biológicas. Laboratório de Estrutura e Função de Proteínas. Juiz de Fora, MG, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Molecular de Tripanossomatídeos. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.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.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-83084https://www.arca.fiocruz.br/bitstream/icict/40905/1/license.txt783568c2893d2e25a99990b126be1772MD51ORIGINALfmicb-11-0043.pdffmicb-11-0043.pdfapplication/pdf2194416https://www.arca.fiocruz.br/bitstream/icict/40905/2/fmicb-11-0043.pdfd5677966db929685dfa43fa437edb874MD52TEXTfmicb-11-0043.pdf.txtfmicb-11-0043.pdf.txtExtracted texttext/plain59622https://www.arca.fiocruz.br/bitstream/icict/40905/3/fmicb-11-0043.pdf.txta150af2f8371469fddf08e857fe60783MD53icict/409052022-06-24 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| dc.title.pt_BR.fl_str_mv |
Knocking down TcNTPDase-1 gene reduces in vitro infectivity of Trypanosoma cruzi |
| title |
Knocking down TcNTPDase-1 gene reduces in vitro infectivity of Trypanosoma cruzi |
| spellingShingle |
Knocking down TcNTPDase-1 gene reduces in vitro infectivity of Trypanosoma cruzi Silva-Gomes, Natália Lins TcNTPDase-1 Gene Knockout Techniques pathogenicity infectivity Enzyme Activation Técnicas de Inactivación de Genes patogenicidad Activación Enzimática Trypanosoma cruzi Técnicas de Inativação de Genes patogenicidade Ativação Enzimática |
| title_short |
Knocking down TcNTPDase-1 gene reduces in vitro infectivity of Trypanosoma cruzi |
| title_full |
Knocking down TcNTPDase-1 gene reduces in vitro infectivity of Trypanosoma cruzi |
| title_fullStr |
Knocking down TcNTPDase-1 gene reduces in vitro infectivity of Trypanosoma cruzi |
| title_full_unstemmed |
Knocking down TcNTPDase-1 gene reduces in vitro infectivity of Trypanosoma cruzi |
| title_sort |
Knocking down TcNTPDase-1 gene reduces in vitro infectivity of Trypanosoma cruzi |
| author |
Silva-Gomes, Natália Lins |
| author_facet |
Silva-Gomes, Natália Lins Rampazzo, Rita de Cássia Pontello Moreira, Claudia Maria do Nascimento Porcino, Gabriane Nascimento Santos, Cyndia Mara Bezerra dos Krieger, Marco Aurélio Vasconcelos, Eveline Gomes Fragoso, Stenio Perdigão Moreira, Otacilio C. |
| author_role |
author |
| author2 |
Rampazzo, Rita de Cássia Pontello Moreira, Claudia Maria do Nascimento Porcino, Gabriane Nascimento Santos, Cyndia Mara Bezerra dos Krieger, Marco Aurélio Vasconcelos, Eveline Gomes Fragoso, Stenio Perdigão Moreira, Otacilio C. |
| author2_role |
author author author author author author author author |
| dc.contributor.author.fl_str_mv |
Silva-Gomes, Natália Lins Rampazzo, Rita de Cássia Pontello Moreira, Claudia Maria do Nascimento Porcino, Gabriane Nascimento Santos, Cyndia Mara Bezerra dos Krieger, Marco Aurélio Vasconcelos, Eveline Gomes Fragoso, Stenio Perdigão Moreira, Otacilio C. |
| dc.subject.other.pt_BR.fl_str_mv |
TcNTPDase-1 |
| topic |
TcNTPDase-1 Gene Knockout Techniques pathogenicity infectivity Enzyme Activation Técnicas de Inactivación de Genes patogenicidad Activación Enzimática Trypanosoma cruzi Técnicas de Inativação de Genes patogenicidade Ativação Enzimática |
| dc.subject.en.pt_BR.fl_str_mv |
Gene Knockout Techniques pathogenicity infectivity Enzyme Activation |
| dc.subject.es.pt_BR.fl_str_mv |
Técnicas de Inactivación de Genes patogenicidad Activación Enzimática |
| dc.subject.decs.pt_BR.fl_str_mv |
Trypanosoma cruzi Técnicas de Inativação de Genes patogenicidade Ativação Enzimática |
| description |
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil. |
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2020 |
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2020-04-21T15:35:39Z |
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2020-04-21T15:35:39Z |
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2020 |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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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. |
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https://www.arca.fiocruz.br/handle/icict/40905 |
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1664-302X |
| dc.identifier.doi.none.fl_str_mv |
10.3389/fmicb.2020.00434 |
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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. 1664-302X 10.3389/fmicb.2020.00434 |
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