title: An efficient low cost method for gene transfer to T lymphocytes.
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.
  
  
  OBJECTIVE
  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.
authors.affiliation: Programa de Carcinogênese Molecular, Coordenação de Pesquisa (CPQ), Instituto Nacional de Câncer (INCA), Rio de Janeiro, Brazil.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Chicaybam
authors.forename: Andressa Laino
authors.initials: AL
authors.lastname: Sodre
authors.forename: Bianca Azevedo
authors.initials: BA
authors.lastname: Curzio
authors.forename: Martin Hernan
authors.initials: MH
authors.lastname: Bonamino
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D018274:Electroporation; D018014:Gene Transfer Techniques; D006801:Humans; D051379:Mice; D013601:T-Lymphocytes / Q000378:metabolism*
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