( the expression of Akt2 with an siRNA ...)RNAi as a Tool for Mammalian Gene Analysis: Applications of siRNAs

the expression of Akt2 with an siRNA, they noted a slight change in cellular glucose regulation. However, when they targeted the two isoforms of the Akt gene (Akt1 and Akt2), they noted a significant change in glucose regulation. Experiments with isoform-specific siRNA ultimately showed that Akt2, and Akt1 to a lesser extent, has an essential role in insulin-stimulated GLUT4 translocation and glucose uptake in two different cell lines, whereas Akt1 and Akt2 contribute equally to insulin-stimulated glycogen synthesis.

Functional Screening
Libraries of siRNAs targeting broad collections of genes will enable screening experiments to tie genes to cellular function. To date, libraries with more than a couple of hundred siRNAs have been limited to a few large research organizations. Recognizing the benefits of siRNA libraries, Ambion is preparing a collection of more than 1800 siRNAs targeting the known human kinases. There have been no published reports on the application of siRNA libraries in screening experiments, but screens in Drosophila and C. elegans using dsRNA libraries exemplify the opportunities.

RNAi libraries targeting more than 10,000 genes have been used in C. elegans to identify genes that regulate fat (9), life expectancy (10), and mutation control (11). A similar RNAi library for Drosophila has been used to identify the genes responsible for regulating the phosphorylation of
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