title: MicroRNA-regulated, systemically delivered rAAV9: a step closer to CNS-restricted transgene expression.
abstract: Recombinant adeno-associated viruses (rAAVs) that can cross the blood-brain-barrier and achieve efficient and stable transvascular gene transfer to the central nervous system (CNS) hold significant promise for treating CNS disorders. However, following intravascular delivery, these vectors also target liver, heart, skeletal muscle, and other tissues, which may cause untoward effects. To circumvent this, we used tissue-specific, endogenous microRNAs (miRNAs) to repress rAAV expression outside the CNS, by engineering perfectly complementary miRNA-binding sites into the rAAV9 genome. This approach allowed simultaneous multi-tissue regulation and CNS-directed stable transgene expression without detectably perturbing the endogenous miRNA pathway. Regulation of rAAV expression by miRNA was primarily via site-specific cleavage of the transgene mRNA, generating specific 5' and 3' mRNA fragments. Our findings promise to facilitate the development of miRNA-regulated rAAV for CNS-targeted gene delivery and other applications.
authors.affiliation: Gene Therapy Center, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Xie
authors.forename: Qing
authors.initials: Q
authors.lastname: Xie
authors.forename: Hongwei
authors.initials: H
authors.lastname: Zhang
authors.forename: Stefan L
authors.initials: SL
authors.lastname: Ameres
authors.forename: Jui-Hung
authors.initials: JH
authors.lastname: Hung
authors.forename: Qin
authors.initials: Q
authors.lastname: Su
authors.forename: Ran
authors.initials: R
authors.lastname: He
authors.forename: Xin
authors.initials: X
authors.lastname: Mu
authors.forename: Seemin
authors.initials: S
authors.lastname: Seher Ahmed
authors.forename: Soyeon
authors.initials: S
authors.lastname: Park
authors.forename: Hiroki
authors.initials: H
authors.lastname: Kato
authors.forename: Chengjian
authors.initials: C
authors.lastname: Li
authors.forename: Christian
authors.initials: C
authors.lastname: Mueller
authors.forename: Craig C
authors.initials: CC
authors.lastname: Mello
authors.forename: Zhiping
authors.initials: Z
authors.lastname: Weng
authors.forename: Terence R
authors.initials: TR
authors.lastname: Flotte
authors.forename: Phillip D
authors.initials: PD
authors.lastname: Zamore
authors.forename: Guangping
authors.initials: G
authors.lastname: Gao
mesh_terms: D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D002490:Central Nervous System / Q000378:metabolism*; D000229:Dependovirus / Q000235:genetics*; D005786:Gene Expression Regulation / Q000235:genetics; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008954:Models, Biological; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D019076:Transgenes / Q000235:genetics
chemical_list: D035683:MicroRNAs