Effective determination of downstreanr signaling events using vector-based RNA interference technologies with modification state-specific antibodies

摘要

The amount of information that RNAi-mediated, targeted gene silencing can provide a researcher is greatly enhanced if it is combined with techniques that seek to determine the contribution the targeted gene has on downstream processes. In this study, it was shown that the vector-based shRNA, pKD-Raf1-v6, was very effective at silencing the Raf1 gene. But what was significant was that the targeted Raf1 reduction by pKD-Raf1-v6 prevented the arsenite and EGF activation of Raf1 downstream effectors Erk1/2 and Rsk1 as demonstrated through the use of phophorylation specific antibodies. These data could not have been generated using the gene chip microarray technology that is often used on samples from targeted, RNAi-mediated gene silencing.