An efficient low cost method for gene transfer to T lymphocytes.

<h4>Unlabelled</h4>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 re...

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Autores principales:	Leonardo Chicaybam, Andressa Laino Sodre, Bianca Azevedo Curzio, Martin Hernan Bonamino
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d9407764592a42e5aa85b95f371bbe51
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spelling	oai:doaj.org-article:d9407764592a42e5aa85b95f371bbe512021-11-18T07:51:52ZAn efficient low cost method for gene transfer to T lymphocytes.1932-620310.1371/journal.pone.0060298https://doaj.org/article/d9407764592a42e5aa85b95f371bbe512013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23555950/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Unlabelled</h4>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.<h4>Aims</h4>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.<h4>Results</h4>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.<h4>Conclusions</h4>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.Leonardo ChicaybamAndressa Laino SodreBianca Azevedo CurzioMartin Hernan BonaminoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 3, p e60298 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Leonardo Chicaybam Andressa Laino Sodre Bianca Azevedo Curzio Martin Hernan Bonamino An efficient low cost method for gene transfer to T lymphocytes.
description	<h4>Unlabelled</h4>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.<h4>Aims</h4>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.<h4>Results</h4>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.<h4>Conclusions</h4>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.
format	article
author	Leonardo Chicaybam Andressa Laino Sodre Bianca Azevedo Curzio Martin Hernan Bonamino
author_facet	Leonardo Chicaybam Andressa Laino Sodre Bianca Azevedo Curzio Martin Hernan Bonamino
author_sort	Leonardo Chicaybam
title	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_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	efficient low cost method for gene transfer to t lymphocytes.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/d9407764592a42e5aa85b95f371bbe51
work_keys_str_mv	AT leonardochicaybam anefficientlowcostmethodforgenetransfertotlymphocytes AT andressalainosodre anefficientlowcostmethodforgenetransfertotlymphocytes AT biancaazevedocurzio anefficientlowcostmethodforgenetransfertotlymphocytes AT martinhernanbonamino anefficientlowcostmethodforgenetransfertotlymphocytes AT leonardochicaybam efficientlowcostmethodforgenetransfertotlymphocytes AT andressalainosodre efficientlowcostmethodforgenetransfertotlymphocytes AT biancaazevedocurzio efficientlowcostmethodforgenetransfertotlymphocytes AT martinhernanbonamino efficientlowcostmethodforgenetransfertotlymphocytes
_version_	1718422882072133632

An efficient low cost method for gene transfer to T lymphocytes.

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