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RNA Interference and Gene Silencing: History and Overview

with these characteristics are effective. The reasons for this are unclear but may be a result of positional effects (46, 48, 49). For current recommendations on designing siRNAs, see "siRNA Design".


RNAi as a Tool for Functional Genomics

Although the history and mechanism of RNAi and PTGS are fascinating, many researchers are most excited about RNAi's potential use as a functional genomics tool. Already RNAi has been used to ascertain the function of many genes in Drosophila, C. elegans, and several species of plants. With the knowledge that RNAi can be induced in mammalian cells by the transfection of siRNAs, many more researchers are beginning to use RNAi as a tool in human, mouse and other mammalian cell culture systems.

In early experiments with mammalian cells, the siRNAs were synthesized chemically (Ambion is one of several companies that offer custom siRNA synthesis). Recently, Ambion introduced a kit (the Silencer siRNA Construction Kit) to produce siRNAs by in vitro transcription, which is a less expensive alternative to chemical synthesis, particularly when multiple different siRNAs need to be synthesized. Once made, the siRNAs are introduced into cells via transient transfection. Due to differences in efficacy, most researchers will synthesize 34 siRNAs to a target gene and perform pilot experiments to determine the most effective one. Transient silencing of more than 90% has been observed wi
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