Although long dsRNAs (several hundred bp) are commonly employed to trigger RNAi in C. elegans or D. melanogaster, these molecules also activate the innate immune system and trigger interferon (IFN) responses in higher organisms. RNAi can be performed in mammalian cells using short RNAs, which generally do not induce IFN responses. Historically, siRNAs have been synthesized as 21mers that bypass the need for Dicer processing by directly mimic the products that are produced by Dicer in vivo.
However, it is now thought that, in addition to being a nuclease, Dicer is also required to introduce the siRNA into RISC and is involved in RISC assembly (Figure 2) [4–6]. IDT DsiRNAs are chemically synthesized 27mer RNA duplexes that are optimized for Dicer processing and show increased potency when compared with 21mer siRNAs [7–8]. Dicer-substrate RNAi methods take advantage of the link between Dicer and RISC loading that occurs when RNAs are processed by Dicer.