A new method for rapidly generating gene-targeting vectors by engineering BACs through homologous recombination in bacteria

Detalhes bibliográficos
Autor(a) principal: Almeida, Vinicius Cotta de
Data de Publicação: 2003
Outros Autores: Schonhoff, Susan, Shibata, Tomoyuki, Leiter, Andrew, Snapper, Scott B.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/41058
Resumo: Massachusetts General Hospital. Gastrointestinal Unit. Center for the Study of Inflammatory Bowel Disease. Boston, MA, USA / Harvard Medical School. Department of Medicine. Boston, MA, USA / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Ultraestrutura e Biologia Celular. Rio de Janeiro, RJ, Brasil.
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spelling Almeida, Vinicius Cotta deSchonhoff, SusanShibata, TomoyukiLeiter, AndrewSnapper, Scott B.2020-05-01T17:11:16Z2020-05-01T17:11:16Z2003ALMEIDA, Vinicius Cotta de et al. A New Method for Rapidly Generating Gene-Targeting Vectors by Engineering BACs Through Homologous Recombination in Bacteria. Genome Research, v. 13, p. 2190-2194, 2003.1088-9051https://www.arca.fiocruz.br/handle/icict/4105810.1101/gr.13565031549-5469engCold Spring Harbor Laboratory PressNovo métodoVetoresCromossomos artificiais bacterianosEscherichia coliRecombinação homólogaNew MethodGenerating Gene-Targeting VectorHomologous RecombinationBacteriaEscherichia coliA new method for rapidly generating gene-targeting vectors by engineering BACs through homologous recombination in bacteriainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleMassachusetts General Hospital. Gastrointestinal Unit. Center for the Study of Inflammatory Bowel Disease. Boston, MA, USA / Harvard Medical School. Department of Medicine. Boston, MA, USA / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Ultraestrutura e Biologia Celular. Rio de Janeiro, RJ, Brasil.New England Medical Center. GRASP Digestive Disease Center. Division of Gastroenterology. Boston, Massachussets, USA / Tufts University School of Medicine, Boston, Massachusetts, USA.Massachusetts General Hospital. Gastrointestinal Unit. Center for the Study of Inflammatory Bowel Disease. Boston, MA, USA / Harvard Medical School. Department of Medicine. Boston, MA, USA .New England Medical Center. GRASP Digestive Disease Center. Division of Gastroenterology. Boston, Massachussets, USA / Tufts University School of Medicine, Boston, Massachusetts, USA.Massachusetts General Hospital. Gastrointestinal Unit. Center for the Study of Inflammatory Bowel Disease. Boston, MA, USA / Harvard Medical School. Department of Medicine. Boston, MA, USA .Generating knockout mice is still an expensive and highly time-consuming process. Target construct generation, the first labor-intensive step in this process, requires the manipulation of large fragments of DNA and numerous, and often cumbersome, cloning steps. Here we show the development of a rapid approach for generating targeting constructs that capitalizes on efficient homologous recombination between linear DNA fragments and circular plasmids in Escherichia coli ("recombineering"), the availability of bacterial artificial chromosomes (BACs), and the accessibility of the sequence of the mouse genome. Employing recombineering, we demonstrate with only 1-2 template plasmids, short homologies (40-50bp) between donor and target DNA, and one subcloning step that we can efficiently manipulate BACs in situ to generate a complicated targeting vector. This procedure avoids the need to construct or screen genomic libraries and permits the generation of most standard, conditional, or knock-in targeting vectors, often within two weeks.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/41058/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALViniciusCottade_Almeida_etal_IOC_2003.pdfViniciusCottade_Almeida_etal_IOC_2003.pdfapplication/pdf282857https://www.arca.fiocruz.br/bitstream/icict/41058/2/ViniciusCottade_Almeida_etal_IOC_2003.pdf59b6c025bc78829d566ea7c333125c09MD52TEXTViniciusCottade_Almeida_etal_IOC_2003.pdf.txtViniciusCottade_Almeida_etal_IOC_2003.pdf.txtExtracted texttext/plain24040https://www.arca.fiocruz.br/bitstream/icict/41058/3/ViniciusCottade_Almeida_etal_IOC_2003.pdf.txtd6701e4738a32a80331c122188c2a6f7MD53icict/410582020-05-02 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dc.title.pt_BR.fl_str_mv A new method for rapidly generating gene-targeting vectors by engineering BACs through homologous recombination in bacteria
title A new method for rapidly generating gene-targeting vectors by engineering BACs through homologous recombination in bacteria
spellingShingle A new method for rapidly generating gene-targeting vectors by engineering BACs through homologous recombination in bacteria
Almeida, Vinicius Cotta de
Novo método
Vetores
Cromossomos artificiais bacterianos
Escherichia coli
Recombinação homóloga
New Method
Generating Gene-Targeting Vector
Homologous Recombination
Bacteria
Escherichia coli
title_short A new method for rapidly generating gene-targeting vectors by engineering BACs through homologous recombination in bacteria
title_full A new method for rapidly generating gene-targeting vectors by engineering BACs through homologous recombination in bacteria
title_fullStr A new method for rapidly generating gene-targeting vectors by engineering BACs through homologous recombination in bacteria
title_full_unstemmed A new method for rapidly generating gene-targeting vectors by engineering BACs through homologous recombination in bacteria
title_sort A new method for rapidly generating gene-targeting vectors by engineering BACs through homologous recombination in bacteria
author Almeida, Vinicius Cotta de
author_facet Almeida, Vinicius Cotta de
Schonhoff, Susan
Shibata, Tomoyuki
Leiter, Andrew
Snapper, Scott B.
author_role author
author2 Schonhoff, Susan
Shibata, Tomoyuki
Leiter, Andrew
Snapper, Scott B.
author2_role author
author
author
author
dc.contributor.author.fl_str_mv Almeida, Vinicius Cotta de
Schonhoff, Susan
Shibata, Tomoyuki
Leiter, Andrew
Snapper, Scott B.
dc.subject.other.pt_BR.fl_str_mv Novo método
Vetores
Cromossomos artificiais bacterianos
Escherichia coli
Recombinação homóloga
topic Novo método
Vetores
Cromossomos artificiais bacterianos
Escherichia coli
Recombinação homóloga
New Method
Generating Gene-Targeting Vector
Homologous Recombination
Bacteria
Escherichia coli
dc.subject.en.pt_BR.fl_str_mv New Method
Generating Gene-Targeting Vector
Homologous Recombination
Bacteria
Escherichia coli
description Massachusetts General Hospital. Gastrointestinal Unit. Center for the Study of Inflammatory Bowel Disease. Boston, MA, USA / Harvard Medical School. Department of Medicine. Boston, MA, USA / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Ultraestrutura e Biologia Celular. Rio de Janeiro, RJ, Brasil.
publishDate 2003
dc.date.issued.fl_str_mv 2003
dc.date.accessioned.fl_str_mv 2020-05-01T17:11:16Z
dc.date.available.fl_str_mv 2020-05-01T17:11:16Z
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 ALMEIDA, Vinicius Cotta de et al. A New Method for Rapidly Generating Gene-Targeting Vectors by Engineering BACs Through Homologous Recombination in Bacteria. Genome Research, v. 13, p. 2190-2194, 2003.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/41058
dc.identifier.issn.pt_BR.fl_str_mv 1088-9051
dc.identifier.doi.none.fl_str_mv 10.1101/gr.1356503
dc.identifier.eissn.none.fl_str_mv 1549-5469
identifier_str_mv ALMEIDA, Vinicius Cotta de et al. A New Method for Rapidly Generating Gene-Targeting Vectors by Engineering BACs Through Homologous Recombination in Bacteria. Genome Research, v. 13, p. 2190-2194, 2003.
1088-9051
10.1101/gr.1356503
1549-5469
url https://www.arca.fiocruz.br/handle/icict/41058
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 Cold Spring Harbor Laboratory Press
publisher.none.fl_str_mv Cold Spring Harbor Laboratory 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/41058/1/license.txt
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