New technologies from siRNA world

摘要

RNA interference (RNAi) has developed into one of the most exciting and important methods for analyzing gene function and metabolic pathways in the past 15 years. The ability to assess gene function via siRNA-mediated methods represents an exciting and valuable tool that has already begun to accelerate critical investigations across abroad range of biomedical and biological research. Recently, widespread, non-specific effects that complicate the interpretation of data generated from siRNA-mediated knockdown studies have been observed. Off-target effects occur -when a siRNA is processed by the RISC and down-regulates unintended targets. As these changes in gene expression can lead to measurable phenotypes (i.e. false positives) it is of great importance to understand the mechanism behind off-targeting so that strategies can be developed to minimize their effects. The research on off-target effects found that a combination of bioinformatics, chemical modifications, and siRNA pooling significantly reduce unintended targeting, leading to the development of new technologies. Characterized by the same path-way of RNAi, microRNAs (miRNAs) have rapidly emerged as an important class of short endogenous RNAs that act as post-transcriptional regulators of gene expression by base-pairing with their target mRNAs. Several technologies for assessing expression profiling and function of microRNAs are available to the research community and presented in this review.