An efficient low cost method for gene transfer to T lymphocytes

Detalhes bibliográficos
Autor(a) principal: Chicaybam, Leonardo
Data de Publicação: 2013
Outros Autores: Sodre, Andressa Laino, Curzio, Bianca Azevedo, Bonamino, Martin Hernan
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/36136
Resumo: Instituto Nacional de Câncer. Coordenação de Pesquisa. Programa de Carcinogênese Molecular. Rio de Janeiro, RJ, Brasil / Fundação Instituto Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Rio de Janeiro, RJ, Brasil.
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spelling Chicaybam, LeonardoSodre, Andressa LainoCurzio, Bianca AzevedoBonamino, Martin Hernan2019-10-03T14:44:34Z2019-10-03T14:44:34Z2013CHICAYBAM, Leonardo et al. An efficient low cost method for gene transfer to T lymphocytes. Plos One, v. 8, n. 3, p. 1-11, Mar. 2013.1932-6203https://www.arca.fiocruz.br/handle/icict/3613610.1371/journal.pone.00602981932-6203engPublic Library of ScienceAn efficient low cost method for gene transfer to T lymphocytesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleInstituto Nacional de Câncer. Coordenação de Pesquisa. Programa de Carcinogênese Molecular. Rio de Janeiro, RJ, Brasil / Fundação Instituto Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Rio de Janeiro, RJ, Brasil.Instituto Nacional de Câncer. Coordenação de Pesquisa. Programa de Carcinogênese Molecular. Rio de Janeiro, RJ, BrasilInstituto Nacional de Câncer. Coordenação de Pesquisa. Programa de Carcinogênese Molecular. Rio de Janeiro, RJ, BrasilInstituto Nacional de Câncer. Coordenação de Pesquisa. Programa de Carcinogênese Molecular. Rio de Janeiro, RJ, BrasilGene transfer to T lymphocytes has historically relied on retro and lentivirus, but recently transposon-based gene transfer is rising as a simpler and straight forward approach to achieve stable transgene expression. Transfer of expression cassettes to T lymphocytes remains challenging, being based mainly on commercial kits. Aims: We herein report a convenient and affordable method based on in house made buffers, generic cuvettes and utilization of the widely available Lonza nucleofector II device to promote efficient gene transfer to T lymphocytes. Results: This approach renders high transgene expression levels in primary human T lymphocytes (mean 45%, 41–59%), the hard to transfect murine T cells (mean 38%, 36–42% for C57/BL6 strain) and human Jurkat T cell line. Cell viability levels after electroporation allowed further manipulations such as in vitro expansion and Chimeric Antigen Receptor (CAR) mediated gain of function for target cell lysis. Conclusions: We describe here an efficient general protocol for electroporation based modification of T lymphocytes. By opening access to this protocol, we expect that efficient gene transfer to T lymphocytes, for transient or stable expression, may be achieved by an increased number of laboratories at lower and affordable costs.T lymphocytesGene transferinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txttext/plain1748https://www.arca.fiocruz.br/bitstream/icict/36136/1/license.txt8a4605be74aa9ea9d79846c1fba20a33MD51ORIGINALve_Chicaybam_Leonardo_etal_INI_2013.pdfve_Chicaybam_Leonardo_etal_INI_2013.pdfapplication/pdf1791199https://www.arca.fiocruz.br/bitstream/icict/36136/2/ve_Chicaybam_Leonardo_etal_INI_2013.pdfeef1f9b801447997c7f187e4cedf3d2cMD52TEXTve_Chicaybam_Leonardo_etal_INI_2013.pdf.txtve_Chicaybam_Leonardo_etal_INI_2013.pdf.txtExtracted texttext/plain54460https://www.arca.fiocruz.br/bitstream/icict/36136/3/ve_Chicaybam_Leonardo_etal_INI_2013.pdf.txt0e7e2a7f75439f287e00a114ebd4e3dbMD53icict/361362019-10-04 02:01:16.898oai:www.arca.fiocruz.br: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Repositório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352019-10-04T05:01:16Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)false
dc.title.pt_BR.fl_str_mv An efficient low cost method for gene transfer to T lymphocytes
title An efficient low cost method for gene transfer to T lymphocytes
spellingShingle An efficient low cost method for gene transfer to T lymphocytes
Chicaybam, Leonardo
T lymphocytes
Gene transfer
title_short An efficient low cost method for gene transfer to T lymphocytes
title_full An efficient low cost method for gene transfer to T lymphocytes
title_fullStr An efficient low cost method for gene transfer to T lymphocytes
title_full_unstemmed An efficient low cost method for gene transfer to T lymphocytes
title_sort An efficient low cost method for gene transfer to T lymphocytes
author Chicaybam, Leonardo
author_facet Chicaybam, Leonardo
Sodre, Andressa Laino
Curzio, Bianca Azevedo
Bonamino, Martin Hernan
author_role author
author2 Sodre, Andressa Laino
Curzio, Bianca Azevedo
Bonamino, Martin Hernan
author2_role author
author
author
dc.contributor.author.fl_str_mv Chicaybam, Leonardo
Sodre, Andressa Laino
Curzio, Bianca Azevedo
Bonamino, Martin Hernan
dc.subject.en.pt_BR.fl_str_mv T lymphocytes
Gene transfer
topic T lymphocytes
Gene transfer
description Instituto Nacional de Câncer. Coordenação de Pesquisa. Programa de Carcinogênese Molecular. Rio de Janeiro, RJ, Brasil / Fundação Instituto Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Rio de Janeiro, RJ, Brasil.
publishDate 2013
dc.date.issued.fl_str_mv 2013
dc.date.accessioned.fl_str_mv 2019-10-03T14:44:34Z
dc.date.available.fl_str_mv 2019-10-03T14:44:34Z
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 CHICAYBAM, Leonardo et al. An efficient low cost method for gene transfer to T lymphocytes. Plos One, v. 8, n. 3, p. 1-11, Mar. 2013.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/36136
dc.identifier.issn.pt_BR.fl_str_mv 1932-6203
dc.identifier.doi.none.fl_str_mv 10.1371/journal.pone.0060298
dc.identifier.eissn.none.fl_str_mv 1932-6203
identifier_str_mv CHICAYBAM, Leonardo et al. An efficient low cost method for gene transfer to T lymphocytes. Plos One, v. 8, n. 3, p. 1-11, Mar. 2013.
1932-6203
10.1371/journal.pone.0060298
url https://www.arca.fiocruz.br/handle/icict/36136
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 Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
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