| Author | Chicaybam, Leonardo | |
| Author | Sodre, Andressa Laino | |
| Author | Curzio, Bianca Azevedo | |
| Author | Bonamino, Martin Hernan | |
| Access date | 2019-10-03T14:44:34Z | |
| Available date | 2019-10-03T14:44:34Z | |
| Document date | 2013 | |
| Citation | 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. | pt_BR |
| ISSN | 1932-6203 | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/36136 | |
| Language | eng | pt_BR |
| Publisher | Public Library of Science | pt_BR |
| Rights | open access | pt_BR |
| Title | An efficient low cost method for gene transfer to T lymphocytes | pt_BR |
| Type | Article | pt_BR |
| DOI | 10.1371/journal.pone.0060298 | |
| Abstract | 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. | pt_BR |
| Affilliation | 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. | pt_BR |
| Affilliation | Instituto Nacional de Câncer. Coordenação de Pesquisa. Programa de Carcinogênese Molecular. Rio de Janeiro, RJ, Brasil | pt_BR |
| Affilliation | Instituto Nacional de Câncer. Coordenação de Pesquisa. Programa de Carcinogênese Molecular. Rio de Janeiro, RJ, Brasil | pt_BR |
| Affilliation | Instituto Nacional de Câncer. Coordenação de Pesquisa. Programa de Carcinogênese Molecular. Rio de Janeiro, RJ, Brasil | pt_BR |
| Subject | T lymphocytes | pt_BR |
| Subject | Gene transfer | pt_BR |
| e-ISSN | 1932-6203 | |
