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FIGURE 2–6. Schematic of the mechanism of RNA interference (RNAi) posttranscriptional knockdown of a gene product.

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FIGURE 2–6. Schematic of the mechanism of RNA interference (RNAi) posttranscriptional knockdown of a gene product.The procedure starts with introduction (transfection, electroporation, or injection) of double-stranded RNA (dsRNA) or small interfering RNA (siRNA) into cells, or expression of small hairpin RNA (shRNA) in cells with vectors encoding shRNAs. The cellular ribonuclease (RNase) Dicer recognizes the long dsRNA molecules and shRNA. Subsequently the dsRNA is cleaved, resulting in 21-nt RNA duplexes, the siRNAs. These siRNA molecules are then incorporated into the RNA-induced silencing complex (RISC) multiprotein complex, where they are unwound by an adenosine triphosphate (ATP)–dependent process, transforming the complex into an active state. Activated RISC uses one strand of the RNA as a bait to bind homologous RNA molecules. The target RNA is cleaved and degraded, resulting in gene silencing.

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