LPG2 Gene Duplication in Leishmania infantum: a Case for CRISPR-Cas9 Gene Editing

Detalhes bibliográficos
Autor(a) principal: Santos, Flávio Henrique Jesus
Data de Publicação: 2020
Outros Autores: Silva, Jéssica Lobo, Ramos, Pablo Ivan Pereira, Descoteaux, Albert, Lima, Jonilson Berlink, Valéria, Matos de Borges, Farias, Leonardo Paiva
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/43366
Resumo: Brazilian National Research Council (CNPq) (Grant Number: 431857/2018-0) and the Fundação de Amparo à Pesquisa do Estado da Bahia (FAPESB) (Grant Number: 04/2015) to VB. VB is a senior investigator funded by CNPq. AD holds the Canada Research Chair in Biology of intracellular parasitism. The fellowships received by FJ-S, JL-S, and part of this study was financed by the program: Coordenação de Aperfeiçoamento de Pessoal deNível Superior— Brasil (CAPES) –Finance Code 001.
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spelling Santos, Flávio Henrique JesusSilva, Jéssica LoboRamos, Pablo Ivan PereiraDescoteaux, AlbertLima, Jonilson BerlinkValéria, Matos de BorgesFarias, Leonardo Paiva2020-09-15T12:13:30Z2020-09-15T12:13:30Z2020SANTOS, Flávio Henrique Jesus et al. LPG2 Gene Duplication in Leishmania infantum: a Case for CRISPR-Cas9 Gene Editing. Frontiers in Cellular and Infection Microbiology, p. 1-11, Aug. 2020.2235-2988https://www.arca.fiocruz.br/handle/icict/4336610.3389/fcimb.2020.00408Brazilian National Research Council (CNPq) (Grant Number: 431857/2018-0) and the Fundação de Amparo à Pesquisa do Estado da Bahia (FAPESB) (Grant Number: 04/2015) to VB. VB is a senior investigator funded by CNPq. AD holds the Canada Research Chair in Biology of intracellular parasitism. The fellowships received by FJ-S, JL-S, and part of this study was financed by the program: Coordenação de Aperfeiçoamento de Pessoal deNível Superior— Brasil (CAPES) –Finance Code 001.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Laboratório de Inflamação e Biomarcadores. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Laboratório de Inflamação e Biomarcadores. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Centro de Integração de Dados e Conhecimentos para Saúde. Salvador, BA, Brasil.Institut National de la Recherche Scientifique. Centre Armand-Frappier Santé Biotechnologie. Laval, QC, Canada.Federal University of Western of Bahia. Center of Biological Sciences and Health. Barreiras, BA, Brazil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Laboratório de Inflamação e Biomarcadores. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Laboratório de Inflamação e Biomarcadores. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina. Salvador, BA, Brasil.On the surface of the Leishmania promastigote, phosphoglycans (PG) such as lipophosphoglycan (LPG), proteophosphoglycan (PPG), free phosphoglycan polymers (PGs), and acid phosphatases (sAP), are dominant and contribute to the invasion and survival of Leishmania within the host cell by modulating macrophage signaling and intracellular trafficking. Phosphoglycan synthesis depends on the Golgi GDP-mannose transporter encoded by the LPG2 gene. Aiming to investigate the role of PG-containing molecules in Leishmania infantum infection process, herein we describe the generation and characterization of L. infantum LPG2-deficient parasites. This gene was unexpectedly identified as duplicated in the L. infantum genome, which impaired gene targeting using the conventional homologous recombination approach. This limitation was circumvented by the use of CRISPR/Cas9 technology. Knockout parasites were selected by agglutination assays using CA7AE antibodies followed by a lectin (RCA 120). Five clones were isolated and molecularly characterized, all revealing the expected edited genome, as well as the complete absence of LPG and PG-containing molecule expression. Finally, the deletion of LPG2 was found to impair the outcome of infection in human neutrophils, as demonstrated by a pronounced reduction (∼83%) in intracellular load compared to wild-type parasite infection. The results obtained herein reinforce the importance of LPG and other PGs as virulence factors in host-parasite interactions.engFrontiers MediaGDP-manoseLeishmania infantumGenesCRISPRGDP-mannose transporterLipophosphoglycanLeishmania infantumGene targetingCRISPR/CAS9LPG2 Gene Duplication in Leishmania infantum: a Case for CRISPR-Cas9 Gene Editinginfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo: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/43366/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALSantos, Flavio Henrique Jesus LPG2 Gene....pdfSantos, Flavio Henrique Jesus LPG2 Gene....pdfapplication/pdf1861232https://www.arca.fiocruz.br/bitstream/icict/43366/2/Santos%2c%20Flavio%20Henrique%20Jesus%20LPG2%20Gene....pdf301bdadb400091d66061c1853f7a43ebMD52TEXTSantos, 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dc.title.pt_BR.fl_str_mv LPG2 Gene Duplication in Leishmania infantum: a Case for CRISPR-Cas9 Gene Editing
title LPG2 Gene Duplication in Leishmania infantum: a Case for CRISPR-Cas9 Gene Editing
spellingShingle LPG2 Gene Duplication in Leishmania infantum: a Case for CRISPR-Cas9 Gene Editing
Santos, Flávio Henrique Jesus
GDP-manose
Leishmania infantum
Genes
CRISPR
GDP-mannose transporter
Lipophosphoglycan
Leishmania infantum
Gene targeting
CRISPR/CAS9
title_short LPG2 Gene Duplication in Leishmania infantum: a Case for CRISPR-Cas9 Gene Editing
title_full LPG2 Gene Duplication in Leishmania infantum: a Case for CRISPR-Cas9 Gene Editing
title_fullStr LPG2 Gene Duplication in Leishmania infantum: a Case for CRISPR-Cas9 Gene Editing
title_full_unstemmed LPG2 Gene Duplication in Leishmania infantum: a Case for CRISPR-Cas9 Gene Editing
title_sort LPG2 Gene Duplication in Leishmania infantum: a Case for CRISPR-Cas9 Gene Editing
author Santos, Flávio Henrique Jesus
author_facet Santos, Flávio Henrique Jesus
Silva, Jéssica Lobo
Ramos, Pablo Ivan Pereira
Descoteaux, Albert
Lima, Jonilson Berlink
Valéria, Matos de Borges
Farias, Leonardo Paiva
author_role author
author2 Silva, Jéssica Lobo
Ramos, Pablo Ivan Pereira
Descoteaux, Albert
Lima, Jonilson Berlink
Valéria, Matos de Borges
Farias, Leonardo Paiva
author2_role author
author
author
author
author
author
dc.contributor.author.fl_str_mv Santos, Flávio Henrique Jesus
Silva, Jéssica Lobo
Ramos, Pablo Ivan Pereira
Descoteaux, Albert
Lima, Jonilson Berlink
Valéria, Matos de Borges
Farias, Leonardo Paiva
dc.subject.other.pt_BR.fl_str_mv GDP-manose
Leishmania infantum
Genes
CRISPR
topic GDP-manose
Leishmania infantum
Genes
CRISPR
GDP-mannose transporter
Lipophosphoglycan
Leishmania infantum
Gene targeting
CRISPR/CAS9
dc.subject.en.pt_BR.fl_str_mv GDP-mannose transporter
Lipophosphoglycan
Leishmania infantum
Gene targeting
CRISPR/CAS9
description Brazilian National Research Council (CNPq) (Grant Number: 431857/2018-0) and the Fundação de Amparo à Pesquisa do Estado da Bahia (FAPESB) (Grant Number: 04/2015) to VB. VB is a senior investigator funded by CNPq. AD holds the Canada Research Chair in Biology of intracellular parasitism. The fellowships received by FJ-S, JL-S, and part of this study was financed by the program: Coordenação de Aperfeiçoamento de Pessoal deNível Superior— Brasil (CAPES) –Finance Code 001.
publishDate 2020
dc.date.accessioned.fl_str_mv 2020-09-15T12:13:30Z
dc.date.available.fl_str_mv 2020-09-15T12:13:30Z
dc.date.issued.fl_str_mv 2020
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 SANTOS, Flávio Henrique Jesus et al. LPG2 Gene Duplication in Leishmania infantum: a Case for CRISPR-Cas9 Gene Editing. Frontiers in Cellular and Infection Microbiology, p. 1-11, Aug. 2020.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/43366
dc.identifier.issn.pt_BR.fl_str_mv 2235-2988
dc.identifier.doi.pt_BR.fl_str_mv 10.3389/fcimb.2020.00408
identifier_str_mv SANTOS, Flávio Henrique Jesus et al. LPG2 Gene Duplication in Leishmania infantum: a Case for CRISPR-Cas9 Gene Editing. Frontiers in Cellular and Infection Microbiology, p. 1-11, Aug. 2020.
2235-2988
10.3389/fcimb.2020.00408
url https://www.arca.fiocruz.br/handle/icict/43366
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Frontiers Media
publisher.none.fl_str_mv Frontiers Media
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
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collection Repositório Institucional da FIOCRUZ (ARCA)
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