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RNAi: A How To for New Users


Reagents Required for RNAi Experiments
The reagents required for inducing and analyzing the RNAi effect are quite simple:

A specific dsRNA that targets a particular gene transcript to induce the RNAi pathway
An efficient dsRNA delivery system
Assays for the RNAi effect
Proper controls


Tools for Non-mammalian RNAi Experiments
In non-mammalian systems such as Caenorhabditis elegans and Drosophila, long dsRNA (typically >200 bp) complementary to the target transcript is used to induce RNAi. The dsRNA can be readily generated by in vitro transcription. The MEGAscript RNAi Kit was developed specifically for this purpose. dsRNA design for inducing RNAi in many non-mammalian systems is straightforwardvirtually any long dsRNA complementary to the targeted transcript will work. Delivery methods are also typically straightforward. For example, for delivery to Drosophila S2 cells, the dsRNA can be added directly to the media used to culture them. dsRNA can also be directly injected into worms or fly embryos. Most researchers use long dsRNAs previously proved to have an easily measured biological effect as a positive control, and dsRNA that does not target any transcript in the organism (e.g., dsRNA targeting luciferase) as a negative control.


Tools for Mammalian RNAi Experiments

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