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An Efficient Low Cost Method for Gene Transfer to T Lymphocytes

Gene 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...

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Detalles Bibliográficos
Autores principales: Chicaybam, Leonardo, Sodre, Andressa Laino, Curzio, Bianca Azevedo, Bonamino, Martin Hernan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3608570/
https://www.ncbi.nlm.nih.gov/pubmed/23555950
http://dx.doi.org/10.1371/journal.pone.0060298
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author Chicaybam, Leonardo
Sodre, Andressa Laino
Curzio, Bianca Azevedo
Bonamino, Martin Hernan
author_facet Chicaybam, Leonardo
Sodre, Andressa Laino
Curzio, Bianca Azevedo
Bonamino, Martin Hernan
author_sort Chicaybam, Leonardo
collection PubMed
description Gene 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.
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spelling pubmed-36085702013-04-03 An Efficient Low Cost Method for Gene Transfer to T Lymphocytes Chicaybam, Leonardo Sodre, Andressa Laino Curzio, Bianca Azevedo Bonamino, Martin Hernan PLoS One Research Article Gene 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. Public Library of Science 2013-03-26 /pmc/articles/PMC3608570/ /pubmed/23555950 http://dx.doi.org/10.1371/journal.pone.0060298 Text en © 2013 Chicaybam et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Chicaybam, Leonardo
Sodre, Andressa Laino
Curzio, Bianca Azevedo
Bonamino, Martin Hernan
An Efficient Low Cost Method for Gene Transfer to T Lymphocytes
title 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_short An Efficient Low Cost Method for Gene Transfer to T Lymphocytes
title_sort efficient low cost method for gene transfer to t lymphocytes
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3608570/
https://www.ncbi.nlm.nih.gov/pubmed/23555950
http://dx.doi.org/10.1371/journal.pone.0060298
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