Efficient Silencing of Gene Expression by an ASON–Bulge–DNAzyme Complex 英文参考文献.docVIP

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Efficient Silencing of Gene Expression by an ASON–Bulge–DNAzyme Complex 英文参考文献.doc

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Efficient Silencing of Gene Expression by an ASON–Bulge–DNAzyme Complex 英文参考文献

EfficientSilencingofGeneExpressionbyanASON– Bulge–DNAzymeComplex JianzhongYi*,ChengqianLiu InstituteofAnimalHusbandryVeterinarySciences,ShanghaiAcademyofAgriculturalSciences,Shanghai,China Abstract Background:DNAzymesareDNAmoleculesthatcandirectlycleavecognatemRNA,andhavebeendevelopedtosilence geneexpressionforresearchandclinicalpurposes.TheadvantageofDNAzymesoverribozymesisthattheyareinexpensive toproduceandexhibitgoodstability.The‘‘10-23DNAenzyme’’iscomposedofacatalyticdomainof15deoxynucleotides, flanked by two substrate-recognition domains of approximately eight nucleotides in each direction, which provides the complementary sequence required for specific binding to RNA substrates. However, these eight nucleotides might not afford sufficient binding energy to hold the RNA substrate along with the DNAzyme, which would interfere with the efficiencyoftheDNAzymeorcausesideeffects,suchasthecleavageofnon-cognatemRNAs. Methodology: In this study, we inserted a nonpairing bulge at the 59 end of the ‘‘10–23 DNA enzyme’’ to enhance its efficiencyandspecificity.Differentsizesofbulgeswereinsertedatdifferentpositionsinthe59endoftheDNAzyme.The non-matching bulge will avoid strong binding between the DNAzyme and target mRNA, which may interfere with the efficiencyoftheDNAzyme. Conclusions:OurnovelDNAzymeconstructscouldefficientlysilencetheexpressionoftargetgenes,provingapowerful toolforgenesilencing.Theresultsshowedthatthesixoligobulgewasthemosteffectivewhenthesixoligobulgewas12– 15bpawayfromthecorecatalyticdomain. Citation: Yi J, Liu C (2011) Efficient Silencing of Gene Expression by an ASON–Bulge–DNAzyme Complex. PLoS ONE 6(4): e18629. doi:10.1371/ journal.pone.0018629 Editor:JosephNajbauer,CityofHopeNationalMedicalCenterandBeckmanResearchInstitute,UnitedStatesofAmerica ReceivedDecember22,2010;AcceptedMarch14,2011;PublishedApril7,2011 Copyright:?2011Yi,Liu.Thisisanopen-accessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense,whichpermitsunrestricted us