TheMechanismofRNAInterference(RNAi)

Longdouble-strandedRNAs(dsRNAs;typically>200nt)canbeusedtosilencetheexpressionoftargetgenesinavarietyoforganismsandcelltypes(e.g.,worms,fruitflies,andplants).Uponintroduction,thelongdsRNAsenteracellularpathwaythatiscommonlyreferredtoastheRNAinterference(RNAi)pathway.First,thedsRNAsgetprocessedinto20-25nucleotide(nt)smallinterferingRNAs(siRNAs)byanRNaseIII-likeenzymecalledDicer(initiationstep).Then,thesiRNAsassembleintoendoribonuclease-containingcomplexesknownasRNA-inducedsilencingcomplexes(RISCs),unwindingintheprocess.ThesiRNAstrandssubsequentlyguidetheRISCstocomplementaryRNAmolecules,wheretheycleaveanddestroythecognateRNA(effecterstep).CleavageofcognateRNAtakesplacenearthemiddleoftheregionboundbythesiRNAstrand.

Inmammaliancells,introductionoflongdsRNA(>30nt)initiatesapotentantiviralresponse,exemplifiedbynonspecificinhibitionofproteinsynthesisandRNAdegradation.Themammalianantiviralresponsecanbebypassed,however,bytheintroductionorexpressionofsiRNAs.

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