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1、METHODSINMOLECULARBIOLOGYMETHODSINMOLECULARBIOLOGYTMTMVolume265RNAInterference,RNAInterference,Editing,Editing,andModificationandModificationMethodsandProtocolsMethodsandProtocolsEditedbyEditedbyJonathaM.GottJonathaM.Gott1RNAInterferenceHistoricalOverviewandSigni
2、ficanceMaryK.MontgomerySummaryIntheearly1990s,attemptstomanipulategeneexpressionbyresearchersworkinginthreedifferentfieldsresultedinunanticipatedgenesilencing.Ratherthanignoringsuchresults,theseresearcherswentontodocumentandfurtherinvestigatethenatureofsuchsilenc
3、ing,whichwasnamed“co-suppression”inplants,“quelling”infungi,and“RNAinterference”(RNAi)innema-todes.Bythelate1990s,itwasdiscoveredthatsilencingcouldbeinitiatedinthisdiversesetoforganismsbyexposingcellstodouble-strandedRNA(dsRNA),whichdirectedthedestructionofmRNAsc
4、ontainingsimilarsequences.Soonafterward,suchdsRNA-mediatedsilencingwasemployedasareversegenetictechniquetoanalyzethefunctionsofspecificgenesinabroadvarietyoforganisms.BiochemicalandgeneticstudiesdesignedtouncoverthecomponentsoftheRNAsilencingmachineryidentifiedac
5、ommoncoreofproteinsthatservetoamplifytheinter-feringRNAsignalanddirectendonucleolyticcleavageoftargetRNAs.AsubsetofsilencingeventsmayalsodirectDNAmethylationoftargetedgenes.RNAsilencingisthoughttohaveevolvedasadefensemechanismtosuppressviralreplicationandtranspos
6、onmobilization.However,additionalfunctionsinvolvingtheRNAimachineryhavebeenuncovered,includingposttranscriptionalregulationofendogenousgenes,andmaintenanceofstructureandfunctionofheterochromatin.Whereasmanyresearchershavefocusedonunderstandingthenaturalbiologi-ca
7、lfunctionsofRNAsilencing,othersaretestingitsutilityinantiviralandcancertherapiesandinotherbiotechnologicalandbiomedicalapplications.KeyWords:RNAsilencing;RNAinterference;cosuppression;quelling;posttranscriptionalgenesilencing;antisenseRNA;double-strandedRNA;dicer
8、;RNA-inducedsilencingcomplex;RNA-directedRNApolymerase.From:MethodsinMolecularBiology,vol.265:RNAInterference,Editing,andModification:MethodsandProtocolsEdited
