Gene silencing of target mRNA by RNA interference (RNAi) has dramatically enhanced the arsenal of genetic tools that can be used to study gene function. RNAi is a process used by plants and invertebrates to specifically inactivate gene function. It has been shown that double-stranded RNA (dsRNA) complementary to particular messenger RNA (mRNA) can specifically inactivate gene function through the RNAi pathway in mammalian systems as well.
Small interfering RNAs (siRNAs) are generated in vivo from long precursor molecules through the action of a specific RNAase III endonuclease called Dicer. Dicer initiates the cleavage of the precursor into ~22 nucleotide (nt) double-stranded duplexes with two nt 3'-overhangs and 5' phosphate termini termed short interfering RNAs (siRNAs). The siRNA is subsequently incorporated into the RNA induced silencing complex (RISC), a protein/RNA complex. The siRNA allows specific recognition of the mRNA target and the endonuclease function of the RISC results in cleavage of mRNA and thus loss of gene expression.
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