Authors
Lequn Li and Vassiliki Boussiotis
Summary
Transfection of primary mouse T cells represents a major breakthrough in addressing research areas as T cell function, activation, and signaling in in vitro systems. In this study, we show transient transfection of genes into naïve mouse T cells using nucleofection, a modified electroporation technique. Using this approach, we knocked down endogenous Smad3 by efficient delivery of Smad3 shRNA into antigen-specific naïve T cells isolated from TCR-transgenic mice. The resultant transfected T cells could be safely adoptively transferred into syngeneic recipients and were fully capable of responding to TCR-mediated activation in vivo. This protocol offers the possibility for rapid functional in vivo studies of targeted genes in primary mouse T cells.Further details
The protocol was published on Protocol Exchange in 2006. To see the entire protocol, click on the source link.Advertisement
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