( P * Al-Khalili et al (2) treated myotu...)RNAi as a Tool for Mammalian Gene Analysis: Applications of siRNAs

P> * Al-Khalili et al (2) treated myotubes with serum and showed that increased glucose uptake correlated with increased cell-surface content of glucose transporter (GLUT1). To confirm that glucose transport depends on GLUT1 expression, cells were treated with GLUT1 siRNA and were shown to have reduced levels of serum-stimulated glucose transport.

* In another report, Chen and Barritt (3) used siRNAs to study the transient receptor potential canonical 1 (TRPC1) gene. The TRPC1 gene was thought to encode a non-selective cation channel activated by depletion of cellular storage and/or an intracellular messenger. When liver cells were treated with the TRPC1 siRNA, they exhibited increased cell volume and decreased inflow of Ca2⁺, Mn²⁺, and ATP in hypotonic solutions supporting the hypothesis.

Target Validation
Much of the excitement surrounding siRNAs has been due to their almost seamless incorporation into the development process for therapeutics. In its simplest form, drug development follows the path of target identification -> target validation -> therapeutic compound development -> compound testing in model systems -> clinical trials. Because they are easy to use and highly specific, siRNAs provide the ultimate tool for validation studies. Reducing the expression of a potential therapeutic target and determining if the desired phenotype results provides confidence that an inhibitor
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