title: Targeting the MicroRNA Passenger Strand for Regulating Therapeutic Transgenes.
abstract: Gene therapy strategies have been developed, which can tissue or disease specifically regulate expression of exogenous transgenes by means of endogenous microRNA (miRNA) activity. However, the use of an endogenous guide strand to regulate an exogenous transgene could affect expression of endogenous miRNA target genes. In this study, we developed a new regulatory system of exogenous transgene expression by targeting the passenger strand. We constructed reporter constructs harboring miRNA-122 guide or passenger target sites with perfect or imperfect complementarity. We observed downregulation of an exogenous transgene harboring the miRNA-122 target sites against either the guide or passenger strand in cells expressing the cognate miRNA or cells stably expressing the miRNA target site. Moreover, the transgene activity as well as the gene expression level increased specifically by intracellular introduction of the antisense RNA against the corresponding strand. Endogenous target gene expression was induced by the transgene construct harboring the miRNA guide strand target sites, but not the passenger strand target sites. Importantly, the therapeutic transgene activity was efficiently regulated by targeting the passenger strand. These results suggested that an approach to passenger strand-regulated expression of therapeutic transgenes could be applied more safely as a therapeutic tool.
authors.affiliation: Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Research Institute of Advanced Omics, Dankook University , Yongin, Republic of Korea.
authors.forename: Sung Jin
authors.initials: SJ
authors.lastname: Kim
authors.affiliation: Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Research Institute of Advanced Omics, Dankook University , Yongin, Republic of Korea.
authors.forename: Chang Ho
authors.initials: CH
authors.lastname: Lee
authors.affiliation: Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Research Institute of Advanced Omics, Dankook University , Yongin, Republic of Korea.
authors.forename: Seong-Wook
authors.initials: SW
authors.lastname: Lee
mesh_terms: D020137:Base Pair Mismatch; D020029:Base Pairing; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic; D017930:Genes, Reporter; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D019169:Jurkat Cells; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D061986:MCF-7 Cells; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D016372:RNA, Antisense / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D019076:Transgenes; D000094704:RNA, Guide, CRISPR-Cas Systems
chemical_list: D008156:Luciferases; D035683:MicroRNAs; D016372:RNA, Antisense; D034741:RNA, Small Interfering; D000094704:RNA, Guide, CRISPR-Cas Systems; C531907:MIRN122 microRNA, human