High-Throughput Functional MicroRNAs Profiling by Recombinant AAV-Based MicroRNA Sensor Arrays 英文参考文献.docVIP

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High-Throughput Functional MicroRNAs Profiling by Recombinant AAV-Based MicroRNA Sensor Arrays 英文参考文献.doc

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High-Throughput Functional MicroRNAs Profiling by Recombinant AAV-Based MicroRNA Sensor Arrays 英文参考文献

High-ThroughputFunctionalMicroRNAsProfilingby RecombinantAAV-BasedMicroRNASensorArrays WenhongTian1,3.,XiaoyanDong2,6.,XuerongLiu3,4,GangWang3,ZheyueDong2,WeiShen2 ,Gang Zheng2,JianxinLu4,JinzhongChen6,YueWang3,ZhijianWu5*,XiaobingWu3* 1College of Life Science, Jilin University, Changchun, Jilin, China, 2Beijing FivePlus Molecular Medicine Institute, Beijing, China, 3State Key Laboratory for Molecular Virology and Genetic Engineering, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China, 4WenzhouMedicalCollege,Wenzhou,Zhejiang,China,5UnitonOcularGeneTherapy,NationalEyeInstitute,NationalInstitutesofHealth,Bethesda,Maryland,United StatesofAmerica,6StateKeyLaboratoryofGeneticEngineering,InstituteofGenetics,SchoolofLifeScience,FudanUniversity,Shanghai,China Abstract Background:microRNAs(miRNAs)aresmallandnon-codingRNAswhichplaycriticalrolesinphysiologicalandpathological processes.AnumberofmethodshavebeenestablishedtodetectandquantifymiRNAexpression.However,methodfor high-throughputmiRNAfunctiondetectionisstilllacking. PrincipalFindings:Wedescribeanadeno-associatedvirus(AAV)vector-basedmicroRNA(miRNA)sensor(Asensor)arrayfor high-throughputfunctionalmiRNAprofiling.EachAsensorcontainsaGaussialuciferase(Gluc)andafireflyluciferase(Fluc) expressioncassettetosensefunctionalmiRNAandtoserveasaninternalcontrolrespectively.Usingthisarray,weacquired functionalprofilesof115miRNAsfor12celllinesandfound‘‘functionalmiRNAsignatures’’forseveralspecificcelllines.The activities of specific miRNAs including the let-7 family, miR-17-92 cluster, miR-221, and miR-222 in HEK 293 cells were compared with theirexpression levels determinedby quantitative reverse transcriptase polymerase chain reaction(QRT- PCR). We also demonstrate two other practical applications of the array, including a comparison of the miRNA activity between HEK293 and HEK293T cells and the ability to monitor miRNA activity changes in K562 cells treat