CRISPR / Cas-9 Vector System: Targets UL-39 and Inhibits HSV-1 Replication in Vitro

Detalhes bibliográficos
Autor(a) principal: Raposo, Jéssica Vasques
Data de Publicação: 2021
Outros Autores: Barros, Luciana Rodrigues Carvalho, Torres, Lauana Ribas, Pinto, Rafaela Barbosa da Silva, Lopes, Amanda de Oliveira, Souza, Elen Mello de, Bonamino, Martin Hernan, Paula, Vanessa Salete de
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/58212
Resumo: Resumo em português - Google tradutor O HSV-1 afeta aproximadamente 67% da população mundial. Aqui, buscamos utilizar o CRISPR/Cas9 sistema com o alvo UL39, essencial para a replicação do vírus. A sequência de sgRNA foi inserida em plasmídeo (PX459-UL39). As células Vero foram transfectadas com PX459-UL39 e a inibição da replicação viral foi avaliada 24 e 48 horas depois usando ensaios de placa e fluorescência e qPCR. Fluorescência análises revelaram a presença de CRISPR/Cas9 anti-HSV-1 dentro das células Vero, e qPCR mostrou que o a carga viral diminuiu em> 95% das células transfectadas com anti-HSV-1 CRISPR/Cas9. Nossos dados demonstram a utilidade do PX459-UL39 para inibir a infecção por HSV-1.
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spelling Raposo, Jéssica VasquesBarros, Luciana Rodrigues CarvalhoTorres, Lauana RibasPinto, Rafaela Barbosa da SilvaLopes, Amanda de OliveiraSouza, Elen Mello deBonamino, Martin HernanPaula, Vanessa Salete de2023-05-08T19:36:30Z2023-05-08T19:36:30Z2021RAPOSO, Jéssica Vasques et al. CRISPR / Cas-9 Vector System: Targets UL-39 and Inhibits HSV-1 Replication in Vitro. Research Square, p. 1 - 10, Oct. 2021.https://www.arca.fiocruz.br/handle/icict/58212Resumo em português - Google tradutor O HSV-1 afeta aproximadamente 67% da população mundial. Aqui, buscamos utilizar o CRISPR/Cas9 sistema com o alvo UL39, essencial para a replicação do vírus. A sequência de sgRNA foi inserida em plasmídeo (PX459-UL39). As células Vero foram transfectadas com PX459-UL39 e a inibição da replicação viral foi avaliada 24 e 48 horas depois usando ensaios de placa e fluorescência e qPCR. Fluorescência análises revelaram a presença de CRISPR/Cas9 anti-HSV-1 dentro das células Vero, e qPCR mostrou que o a carga viral diminuiu em> 95% das células transfectadas com anti-HSV-1 CRISPR/Cas9. Nossos dados demonstram a utilidade do PX459-UL39 para inibir a infecção por HSV-1.HSV-1 affects approximately 67% of the world population. Here, we sought to use the CRISPR / Cas9 system with the UL39 target, essential for virus replication. The sgRNA sequence was inserted into plasmid (PX459-UL39). Vero cells were transfected with PX459-UL39, and inhibition of viral replication was assessed 24 and 48 hours later using plaque assays and fluorescence and qPCR. Fluorescence analyzes revealed the presence of anti-HSV-1 CRISPR/Cas9 within Vero cells, and qPCR showed that the viral load decreased by> 95% of cells transfected with anti-HSV-1 CRISPR / Cas9. Our data demonstrate the usefulness of the PX459-UL39 to inhibit HSV-1 infection.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.INCA: Instituto Nacional de Câncer,Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.INCA: Instituto Nacional de Cancer. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, BrasengResearch SquareHSV-1;sgRNAFluorescenceCRISPR/Cas9CRISPRCas-9 Vector System:Targets UL-39Inhibits HSV-1 Replication in VitroCRISPR / Cas-9 Vector System: Targets UL-39 and Inhibits HSV-1 Replication in Vitroinfo: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/58212/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALJessicaRaposo_etal_IOC_preprint_2023.pdfJessicaRaposo_etal_IOC_preprint_2023.pdfapplication/pdf1286303https://www.arca.fiocruz.br/bitstream/icict/58212/2/JessicaRaposo_etal_IOC_preprint_2023.pdfc2c630d85ce0ccd9f6cbca7d5310235fMD52icict/582122023-05-08 16:36:31.951oai:www.arca.fiocruz.br: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ório 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dc.title.en_US.fl_str_mv CRISPR / Cas-9 Vector System: Targets UL-39 and Inhibits HSV-1 Replication in Vitro
title CRISPR / Cas-9 Vector System: Targets UL-39 and Inhibits HSV-1 Replication in Vitro
spellingShingle CRISPR / Cas-9 Vector System: Targets UL-39 and Inhibits HSV-1 Replication in Vitro
Raposo, Jéssica Vasques
HSV-1;
sgRNA
Fluorescence
CRISPR/Cas9
CRISPR
Cas-9 Vector System:
Targets UL-39
Inhibits HSV-1 Replication in Vitro
title_short CRISPR / Cas-9 Vector System: Targets UL-39 and Inhibits HSV-1 Replication in Vitro
title_full CRISPR / Cas-9 Vector System: Targets UL-39 and Inhibits HSV-1 Replication in Vitro
title_fullStr CRISPR / Cas-9 Vector System: Targets UL-39 and Inhibits HSV-1 Replication in Vitro
title_full_unstemmed CRISPR / Cas-9 Vector System: Targets UL-39 and Inhibits HSV-1 Replication in Vitro
title_sort CRISPR / Cas-9 Vector System: Targets UL-39 and Inhibits HSV-1 Replication in Vitro
author Raposo, Jéssica Vasques
author_facet Raposo, Jéssica Vasques
Barros, Luciana Rodrigues Carvalho
Torres, Lauana Ribas
Pinto, Rafaela Barbosa da Silva
Lopes, Amanda de Oliveira
Souza, Elen Mello de
Bonamino, Martin Hernan
Paula, Vanessa Salete de
author_role author
author2 Barros, Luciana Rodrigues Carvalho
Torres, Lauana Ribas
Pinto, Rafaela Barbosa da Silva
Lopes, Amanda de Oliveira
Souza, Elen Mello de
Bonamino, Martin Hernan
Paula, Vanessa Salete de
author2_role author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Raposo, Jéssica Vasques
Barros, Luciana Rodrigues Carvalho
Torres, Lauana Ribas
Pinto, Rafaela Barbosa da Silva
Lopes, Amanda de Oliveira
Souza, Elen Mello de
Bonamino, Martin Hernan
Paula, Vanessa Salete de
dc.subject.other.en_US.fl_str_mv HSV-1;
sgRNA
Fluorescence
CRISPR/Cas9
topic HSV-1;
sgRNA
Fluorescence
CRISPR/Cas9
CRISPR
Cas-9 Vector System:
Targets UL-39
Inhibits HSV-1 Replication in Vitro
dc.subject.en.en_US.fl_str_mv CRISPR
Cas-9 Vector System:
Targets UL-39
Inhibits HSV-1 Replication in Vitro
description Resumo em português - Google tradutor O HSV-1 afeta aproximadamente 67% da população mundial. Aqui, buscamos utilizar o CRISPR/Cas9 sistema com o alvo UL39, essencial para a replicação do vírus. A sequência de sgRNA foi inserida em plasmídeo (PX459-UL39). As células Vero foram transfectadas com PX459-UL39 e a inibição da replicação viral foi avaliada 24 e 48 horas depois usando ensaios de placa e fluorescência e qPCR. Fluorescência análises revelaram a presença de CRISPR/Cas9 anti-HSV-1 dentro das células Vero, e qPCR mostrou que o a carga viral diminuiu em> 95% das células transfectadas com anti-HSV-1 CRISPR/Cas9. Nossos dados demonstram a utilidade do PX459-UL39 para inibir a infecção por HSV-1.
publishDate 2021
dc.date.issued.fl_str_mv 2021
dc.date.accessioned.fl_str_mv 2023-05-08T19:36:30Z
dc.date.available.fl_str_mv 2023-05-08T19:36:30Z
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 RAPOSO, Jéssica Vasques et al. CRISPR / Cas-9 Vector System: Targets UL-39 and Inhibits HSV-1 Replication in Vitro. Research Square, p. 1 - 10, Oct. 2021.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/58212
identifier_str_mv RAPOSO, Jéssica Vasques et al. CRISPR / Cas-9 Vector System: Targets UL-39 and Inhibits HSV-1 Replication in Vitro. Research Square, p. 1 - 10, Oct. 2021.
url https://www.arca.fiocruz.br/handle/icict/58212
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 Research Square
publisher.none.fl_str_mv Research Square
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/58212/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/58212/2/JessicaRaposo_etal_IOC_preprint_2023.pdf
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