CREditing: a tool for gene tuning in Trypanosoma cruzi

Detalhes bibliográficos
Autor(a) principal: Pacheco-Lugo, Lisandro A.
Data de Publicação: 2020
Outros Autores: Sáenz-García, José L., Díaz-Olmos, Yirys, Netto-Costa, Rodrigo, Brant, Rodrigo S.C., DaRocha, Wanderson D.
Tipo de documento: Artigo
Idioma: por
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/57623
https://doi.org/10.1016/j.ijpara.2020.06.010
Resumo: Universidade Federal do Paraná. Laboratório de Genômica Funcional de Parasitos. Curitiba, PR, Brasil. / Universidad Simón Bolívar. Facultad de Ciencias Básicas y Biomédicas. Barranquilla, Colombia.
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spelling Pacheco-Lugo, Lisandro A.Sáenz-García, José L.Díaz-Olmos, YirysNetto-Costa, RodrigoBrant, Rodrigo S.C.DaRocha, Wanderson D.2023-03-30T12:03:44Z2023-03-30T12:03:44Z2020PACHECO-LUGO, Lisandro A. et al. CREditing: a tool for gene tuning in Trypanosoma cruzi. International Journal for Parasitology, v. 50, p. 1067–1077, 2020.0020-7519https://www.arca.fiocruz.br/handle/icict/57623https://doi.org/10.1016/j.ijpara.2020.06.010porElsevierTrypanosoma cruziCRECRE-loxRecombinasesEnzymesGene DeletionGene activationRecombinasasEnzimasEliminación de GenRecombinasesEnzymesDélétion de gèneRecombinasesEnzimasDeleção de GenesAtivação GênicaCREditing: a tool for gene tuning in Trypanosoma cruziinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleUniversidade Federal do Paraná. Laboratório de Genômica Funcional de Parasitos. Curitiba, PR, Brasil. / Universidad Simón Bolívar. Facultad de Ciencias Básicas y Biomédicas. Barranquilla, Colombia.Universidade Federal do Paraná. Laboratório de Genômica Funcional de Parasitos. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil. / Universidad del Norte. Facultad de Ciencias de la Salud. Barranquilla, Colombia.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Universidade Federal do Paraná. Laboratório de Genômica Funcional de Parasitos. Curitiba, PR, Brasil.Universidade Federal do Paraná. Laboratório de Genômica Funcional de Parasitos. Curitiba, PR, Brasil.The genetic manipulation of Trypanosoma cruzi continues to be a challenge, mainly due to the lack of available and efficient molecular tools. The CRE-lox recombination system is a site-specific recombinase technology, widely used method of achieving conditional targeted deletions, inversions, insertions, gene activation, translocation, and other modifications in chromosomal or episomal DNA. In the present study, the CRE-lox system was adapted to expand the current genetic toolbox for this hard-to-manipulate parasite. For this, evaluations of whether direct protein delivery of CRE recombinase through electroporation could improve CRE-mediated recombination in T. cruzi were performed. CRE recombinase was fused to the C-terminus of T. cruzi histone H2B, which carries the nuclear localization signal and is expressed in the prokaryotic system. The fusion protein was affinity purified and directly introduced into epimastigotes and tissue culture-derived trypomastigotes. This enabled the control of gene expression as demonstrated by turning on a tandem dimer fluorescent protein reporter gene that had been previously transfected into parasites, achieving CRE-mediated recombination in up to 85% of parasites. This system was further tested for its ability to turn off gene expression, remove selectable markers integrated into the genome, and conditionally knock down the nitroreductase gene, which is involved in drug resistance. Additionally, CREditing also enabled the control of gene expression in tissue culture trypomastigotes, which are more difficult to transfect than epimastigotes. The considerable advances in genomic manipulation of T. cruzi shown in this study can be used by others to aid in the greater understanding of this parasite through gain- or loss-of-function approaches.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/57623/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALCREditing--a-tool-for-gene-tuning-in-Trypa_Netto-Rodrigo.pdfCREditing--a-tool-for-gene-tuning-in-Trypa_Netto-Rodrigo.pdfapplication/pdf2255709https://www.arca.fiocruz.br/bitstream/icict/57623/2/CREditing--a-tool-for-gene-tuning-in-Trypa_Netto-Rodrigo.pdf62f95adddb955145acfae0878156cf3cMD52icict/576232023-03-30 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dc.title.en_US.fl_str_mv CREditing: a tool for gene tuning in Trypanosoma cruzi
title CREditing: a tool for gene tuning in Trypanosoma cruzi
spellingShingle CREditing: a tool for gene tuning in Trypanosoma cruzi
Pacheco-Lugo, Lisandro A.
Trypanosoma cruzi
CRE
CRE-lox
Recombinases
Enzymes
Gene Deletion
Gene activation
Recombinasas
Enzimas
Eliminación de Gen
Recombinases
Enzymes
Délétion de gène
Recombinases
Enzimas
Deleção de Genes
Ativação Gênica
title_short CREditing: a tool for gene tuning in Trypanosoma cruzi
title_full CREditing: a tool for gene tuning in Trypanosoma cruzi
title_fullStr CREditing: a tool for gene tuning in Trypanosoma cruzi
title_full_unstemmed CREditing: a tool for gene tuning in Trypanosoma cruzi
title_sort CREditing: a tool for gene tuning in Trypanosoma cruzi
author Pacheco-Lugo, Lisandro A.
author_facet Pacheco-Lugo, Lisandro A.
Sáenz-García, José L.
Díaz-Olmos, Yirys
Netto-Costa, Rodrigo
Brant, Rodrigo S.C.
DaRocha, Wanderson D.
author_role author
author2 Sáenz-García, José L.
Díaz-Olmos, Yirys
Netto-Costa, Rodrigo
Brant, Rodrigo S.C.
DaRocha, Wanderson D.
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Pacheco-Lugo, Lisandro A.
Sáenz-García, José L.
Díaz-Olmos, Yirys
Netto-Costa, Rodrigo
Brant, Rodrigo S.C.
DaRocha, Wanderson D.
dc.subject.other.en_US.fl_str_mv Trypanosoma cruzi
CRE
topic Trypanosoma cruzi
CRE
CRE-lox
Recombinases
Enzymes
Gene Deletion
Gene activation
Recombinasas
Enzimas
Eliminación de Gen
Recombinases
Enzymes
Délétion de gène
Recombinases
Enzimas
Deleção de Genes
Ativação Gênica
dc.subject.en.en_US.fl_str_mv CRE-lox
Recombinases
Enzymes
Gene Deletion
Gene activation
dc.subject.es.en_US.fl_str_mv Recombinasas
Enzimas
Eliminación de Gen
dc.subject.fr.en_US.fl_str_mv Recombinases
Enzymes
Délétion de gène
dc.subject.decs.en_US.fl_str_mv Recombinases
Enzimas
Deleção de Genes
Ativação Gênica
description Universidade Federal do Paraná. Laboratório de Genômica Funcional de Parasitos. Curitiba, PR, Brasil. / Universidad Simón Bolívar. Facultad de Ciencias Básicas y Biomédicas. Barranquilla, Colombia.
publishDate 2020
dc.date.issued.fl_str_mv 2020
dc.date.accessioned.fl_str_mv 2023-03-30T12:03:44Z
dc.date.available.fl_str_mv 2023-03-30T12:03:44Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv PACHECO-LUGO, Lisandro A. et al. CREditing: a tool for gene tuning in Trypanosoma cruzi. International Journal for Parasitology, v. 50, p. 1067–1077, 2020.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/57623
dc.identifier.issn.en_US.fl_str_mv 0020-7519
dc.identifier.doi.none.fl_str_mv https://doi.org/10.1016/j.ijpara.2020.06.010
identifier_str_mv PACHECO-LUGO, Lisandro A. et al. CREditing: a tool for gene tuning in Trypanosoma cruzi. International Journal for Parasitology, v. 50, p. 1067–1077, 2020.
0020-7519
url https://www.arca.fiocruz.br/handle/icict/57623
https://doi.org/10.1016/j.ijpara.2020.06.010
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/57623/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/57623/2/CREditing--a-tool-for-gene-tuning-in-Trypa_Netto-Rodrigo.pdf
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repository.name.fl_str_mv Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)
repository.mail.fl_str_mv repositorio.arca@fiocruz.br
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