Drs. Qi Chen and David Schubert at the Salk Institute recently used in vitro transcribed siRNAs, prepared with Ambion's Silencer siRNA Construction Kit, to study two genes that are involved in nerve cell adhesion. These researchers sought to determine whether siRNA-mediated silencing of Gene 1 would affect the protein expression levels of a putative downstream gene, Gene 2. To this end, the researchers prepared six siRNAs by in vitro transcription that targeted Gene 1. These siRNAs were transfected individually into the nerve cell line B103, and the protein expression levels of both genes were monitored by Western blot 96 hours post-transfection.
One of the six gene specific siRNAs effectively reduced
Gene 1 protein levels (Figure 1A). Interestingly, the same siRNA also
reduced the protein levels of Gene 2, but not those of the -actin control
(Figure 1B and C). These results suggested that Gene 1 does indeed positive