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    Validating rnai targets

    Human filarial nematodes cause river blindness and lymphatic filariasis, both of which are diseases that produce considerable morbidity.Control of these diseases relies on drug treatments that are ineffective against macrofilariae and are threatened by the development of resistance.Genome-wide screens using RNA interference have proven powerful in elucidating components of functionally related pathways and have therefore become integral for the development of new and improved therapeutic targets. This article provides an overview of many of the systems biology approaches taken, using RNA interference, in order to demonstrate how it may be used today for drug discovery and tomorrow as a targeted therapy. Initially undertaken in model organisms such as , functional genomics has now moved into the realm of mammalian cells both in vitro and in vivo due to the development of RNA interference. RNA interference is a conserved biological process that has evolved to specifically and efficiently silence genes.

    validating rnai targets-6validating rnai targets-67

    This study developed a new generation lipid–polymer hybrid nanoparticle platform for effective systemic delivery of small interfering RNA (si RNA) to tumors, which represents a challenging hurdle for the widespread application of RNA interference (RNAi) in cancer research and therapy.

    Two Applied Biosystems scientists have streamlined this step more than 10-fold, performing real-time RT-PCR on cells treated with over 1500 si RNAs targeting more than 500 genes within a single week. For large-scale si RNA screens, the validation process can be very time-consuming.

    This article describes a streamlined workflow for measuring si RNA-induced knockdown of m RNA targets using real-time RT-PCR.

    By eliminating the laborious RNA isolation step and reducing the number of plates and tips needed to process the samples, this streamlined workflow enabled the analysis of target knockdown by over 1500 si RNAs in a single week with minimal hands-on time.

    The workflow employs the Taq Man® Gene Expression Cells-to-CT™ Kit and Taq Man Gene Expression Assays to measure the effect of each si RNA on target gene expression.

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