Disturbance of the microRNA pathway by commonly used lentiviral shRNA libraries limits the application for screening host factors involved in hepatitis C virus infection

摘要

RNAinterference(RNAi)iswidelyusedasascreeningtoolfortheidentificationofhostgenesinvolvedinviralinfection.DuetothelimitationofrawsmallinterferingRNA(siRNA),wetestedtwocommonlyusedshorthairpinRNA(shRNA)lentivirallibrariestoidentifyhostfactorsinvolvedinhepatitisCvirus(HCV)infection.ItwasfoundthattheseshRNAlibraryvectorscausednon‐specificdisturbanceofHCVreplicationthatwasnotduetotoxicityorinterferonresponse,butrelatedtothehighshRNAlevelsdisturbingtheendogenousmicroRNAbiogenesis.ThehighshRNAlevelsachievedwiththesevectorsreducedthelevelsofmaturemicroRNAs,includingmiR‐122knowntopromoteHCVreplication.Ourfindingsextendthecautionofpotentialoff‐targeteffectsoflentiviralshRNAlibrarieswhichappearunsuitabletoscreenmicroRNAregulatedphenotypes,suchasHCVreplication...