Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi

Detalhes bibliográficos
Autor(a) principal: Picchi-Constante, Gisele Fernanda Assine
Data de Publicação: 2022
Outros Autores: Hiraiwa, Priscila Mazzocchi, Marek, Martin, Rogerio, Vanessa Zulkievicz, Guerra-Slompo, Eloise Pavão, Romier, Christophe, Zanchin, Nilson Ivo Tonin
Tipo de documento: Artigo
Idioma: por
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/52921
Resumo: Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
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spelling Picchi-Constante, Gisele Fernanda AssineHiraiwa, Priscila MazzocchiMarek, MartinRogerio, Vanessa ZulkieviczGuerra-Slompo, Eloise PavãoRomier, ChristopheZanchin, Nilson Ivo Tonin2022-05-26T12:04:46Z2022-05-26T12:04:46Z2022PICCHI-CONSTANTE, Gisele Fernanda Assine et al. Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi. Star Protocols, v. 3, n. 101324, p. 1–26, 2022.2666-1667https://www.arca.fiocruz.br/handle/icict/5292110.1016/j.xpro.2022.101324porCell PressTrypanosoma cruziGene ExpressionCRISPR-Associated Protein 9ProtocolsGene EditingExpresión GénicaProteína 9 Asociada a CRISPREdición GénicaExpression des gènesProtéine-9 associée à CRISPRProtocolesÉdition de gèneExpressão GênicaProteína 9 Associada à CRISPRProtocolosEdição de GenesEfficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruziinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Université de Strasbourg, CNRS, INSERM. Institut de Génétique et de Biologie Moléculaire et Cellulaire. Illkirch, France. / Université de Strasbourg, CNRS, INSERM. Institut de Génétique et de Biologie Moléculaire et Cellulaire. Department of Integrated Structural Biology. Illkirch, France. / Loschmidt Laboratories. Department of Experimental Biology & RECETOX. Faculty of Science. Masaryk University. Brno, Czech Republic.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil. / Universidade Federal do Paraná. Programa de Pós-Graduação em Biologia Celular e Molecular. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Université de Strasbourg, CNRS, INSERM. Institut de Génétique et de Biologie Moléculaire et Cellulaire. Illkirch, France. / Université de Strasbourg, CNRS, INSERM. Institut de Génétique et de Biologie Moléculaire et Cellulaire. Department of Integrated Structural Biology. Illkirch, France.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.This protocol outlines a new genetic complementation strategy to investigate gene function in Trypanosoma cruzi, the parasite causing Chagas disease. We combine CRISPR-Cas9 technology with recombination of variants of the target gene containing the desired mutations that are resistant to Cas9-cleavage, which enables detailed investigation of protein function. This experimental strategy overcomes some of the limitations associated with gene knockouts in T. cruzi. For complete details on the use and execution of this protocol, please refer to Marek et al. (2021).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/52921/1/license.txt783568c2893d2e25a99990b126be1772MD51ORIGINALPicchi-Constante_Gisele-EfficientCRISPROK.pdfPicchi-Constante_Gisele-EfficientCRISPROK.pdfapplication/pdf4324224https://www.arca.fiocruz.br/bitstream/icict/52921/2/Picchi-Constante_Gisele-EfficientCRISPROK.pdfd7e19d31b75871b8f903e1edd02b5e91MD52icict/529212022-05-26 09:04:46.926oai:www.arca.fiocruz.br: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ório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352022-05-26T12:04:46Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)false
dc.title.pt_BR.fl_str_mv Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi
title Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi
spellingShingle Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi
Picchi-Constante, Gisele Fernanda Assine
Trypanosoma cruzi
Gene Expression
CRISPR-Associated Protein 9
Protocols
Gene Editing
Expresión Génica
Proteína 9 Asociada a CRISPR
Edición Génica
Expression des gènes
Protéine-9 associée à CRISPR
Protocoles
Édition de gène
Expressão Gênica
Proteína 9 Associada à CRISPR
Protocolos
Edição de Genes
title_short Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi
title_full Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi
title_fullStr Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi
title_full_unstemmed Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi
title_sort Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi
author Picchi-Constante, Gisele Fernanda Assine
author_facet Picchi-Constante, Gisele Fernanda Assine
Hiraiwa, Priscila Mazzocchi
Marek, Martin
Rogerio, Vanessa Zulkievicz
Guerra-Slompo, Eloise Pavão
Romier, Christophe
Zanchin, Nilson Ivo Tonin
author_role author
author2 Hiraiwa, Priscila Mazzocchi
Marek, Martin
Rogerio, Vanessa Zulkievicz
Guerra-Slompo, Eloise Pavão
Romier, Christophe
Zanchin, Nilson Ivo Tonin
author2_role author
author
author
author
author
author
dc.contributor.author.fl_str_mv Picchi-Constante, Gisele Fernanda Assine
Hiraiwa, Priscila Mazzocchi
Marek, Martin
Rogerio, Vanessa Zulkievicz
Guerra-Slompo, Eloise Pavão
Romier, Christophe
Zanchin, Nilson Ivo Tonin
dc.subject.other.pt_BR.fl_str_mv Trypanosoma cruzi
topic Trypanosoma cruzi
Gene Expression
CRISPR-Associated Protein 9
Protocols
Gene Editing
Expresión Génica
Proteína 9 Asociada a CRISPR
Edición Génica
Expression des gènes
Protéine-9 associée à CRISPR
Protocoles
Édition de gène
Expressão Gênica
Proteína 9 Associada à CRISPR
Protocolos
Edição de Genes
dc.subject.en.pt_BR.fl_str_mv Gene Expression
CRISPR-Associated Protein 9
Protocols
Gene Editing
dc.subject.es.pt_BR.fl_str_mv Expresión Génica
Proteína 9 Asociada a CRISPR
Edición Génica
dc.subject.fr.pt_BR.fl_str_mv Expression des gènes
Protéine-9 associée à CRISPR
Protocoles
Édition de gène
dc.subject.decs.pt_BR.fl_str_mv Expressão Gênica
Proteína 9 Associada à CRISPR
Protocolos
Edição de Genes
description Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
publishDate 2022
dc.date.accessioned.fl_str_mv 2022-05-26T12:04:46Z
dc.date.available.fl_str_mv 2022-05-26T12:04:46Z
dc.date.issued.fl_str_mv 2022
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 PICCHI-CONSTANTE, Gisele Fernanda Assine et al. Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi. Star Protocols, v. 3, n. 101324, p. 1–26, 2022.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/52921
dc.identifier.issn.pt_BR.fl_str_mv 2666-1667
dc.identifier.doi.none.fl_str_mv 10.1016/j.xpro.2022.101324
identifier_str_mv PICCHI-CONSTANTE, Gisele Fernanda Assine et al. Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi. Star Protocols, v. 3, n. 101324, p. 1–26, 2022.
2666-1667
10.1016/j.xpro.2022.101324
url https://www.arca.fiocruz.br/handle/icict/52921
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 Cell Press
publisher.none.fl_str_mv Cell Press
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/52921/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/52921/2/Picchi-Constante_Gisele-EfficientCRISPROK.pdf
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repository.name.fl_str_mv Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)
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