title: RNA interference-mediated gene silencing in murine T cells: in vitro and in vivo validation of proinflammatory target genes.
abstract: BACKGROUND
  T cells play a central role in many inflammatory diseases, hence the identification and validation of T cell-specific target genes will increase the understanding of T cell function in pathologic inflammatory situations. RNA interference (RNAi), with its ability to induce specific gene silencing in mammalian cells, represents a powerful technology to investigate and validate the function of pharmaceutical target genes in vitro and in vivo. The aim of the present study was to systematically explore RNAi-mediated gene-silencing of known T cell-specific model signaling molecules in primary murine T cells in vitro and in vivo.
  
  
  RESULTS
  We demonstrate that siRNA delivery and subsequent silencing of T cell specific genes is substantially increased, if murine T cells were activated prior siRNA transfection. Silencing of ZAP70, p56Lck as well as PLC-gamma1 protein expression resulted in impaired function of T cells in vitro. Furthermore, delayed type hypersensitivity (DTH) was ameliorated in vivo after adoptive transfer of ZAP70-silenced T cells.
  
  
  CONCLUSIONS
  The combination of RNAi-mediated gene silencing and adoptive transfer of gene-silenced T cells, thus, may allow the identification and analysis of T cell-specific targets for therapeutic intervention. Additionally, this model system may represent an alternative to conventional time consuming and cost intensive gene targeting approaches.
authors.affiliation: Common Mechanism Research, Bayer Schering Pharma AG, Muellerstrasse 178, 13342, Berlin, Germany. arnevon.bonin@bayerhealthcare.com.
authors.forename: Tatjana C
authors.initials: TC
authors.lastname: Gust
authors.forename: Luisa
authors.initials: L
authors.lastname: Neubrandt
authors.forename: Claudia
authors.initials: C
authors.lastname: Merz
authors.forename: Khusru
authors.initials: K
authors.lastname: Asadullah
authors.forename: Ulrich
authors.initials: U
authors.lastname: Zügel
authors.forename: Arne
authors.initials: A
authors.lastname: von Bonin