Biochemical Dissection of RNA Silencing in Plants
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Although RNA silencing was first discovered in plants, thus far it has been studied biochemically only in animals, where it
            is known as RNA interference (RNAi). In animals, two components of the RNAi pathway have been identified: Dicer, a multidomain
            RNase III that converts long double-stranded RNA (dsRNA) into small interfering RNA (siRNA) and the RNA-induced silencing
            complex (RISC), as siRNA-containing protein–RNA complex that targets complementary mRNA for destruction. We have developed
            methods for the biochemical dissection of plant RNA silencing. In this chapter, we describe in detail how to use wheat germ
            extract to study two distinct Dicer-like activities, RNA-dependent RNA polymerase (RdRP), and endogenous microRNA-programmed
            RISC activities. These comprehensive protocols should prove useful in the further dissection of the plant RNA silencing pathway,
            as well as for the validation of the predicted targets of endogenous plant microRNAs.
         
      









