Please use this identifier to cite or link to this item: https://doi.org/10.1261/rna.031831.111
DC FieldValue
dc.titlemiR-sens - A retroviral dual-luciferase reporter to detect microRNA activity in primary cells
dc.contributor.authorBeillard, E.
dc.contributor.authorOng, S.C.
dc.contributor.authorGiannakakis, A.
dc.contributor.authorGuccione, E.
dc.contributor.authorVardy, L.A.
dc.contributor.authorVoorhoeve, P.M.
dc.date.accessioned2014-11-26T08:29:26Z
dc.date.available2014-11-26T08:29:26Z
dc.date.issued2012-05
dc.identifier.citationBeillard, E., Ong, S.C., Giannakakis, A., Guccione, E., Vardy, L.A., Voorhoeve, P.M. (2012-05). miR-sens - A retroviral dual-luciferase reporter to detect microRNA activity in primary cells. RNA 18 (5) : 1091-1100. ScholarBank@NUS Repository. https://doi.org/10.1261/rna.031831.111
dc.identifier.issn13558382
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/110178
dc.description.abstractMicroRNA-mRNA interactions are commonly validated and deconstructed in cell lines transfected with luciferase reporters. However, due to cell type-specific variations in microRNA or RNA-binding protein abundance, such assays may not reliably reflect microRNA activity in other cell types that are less easily transfected. In order to measure miRNA activity in primary cells, we constructed miR-Sens, a MSCV-based retroviral vector that encodes both a Renilla luciferase reporter gene controlled by microRNA binding sites in its 3′ UTR and a Firefly luciferase normalization gene. miR-Sens sensors can be efficiently transduced in primary cells such as human fibroblasts and mammary epithelial cells, and allow the detection of overexpressed and, more importantly, endogenous microRNAs. Notably, we find that the relative luciferase activity is correlated to the miRNA expression, allowing quantitative measurement of microRNA activity. We have subsequently validated the miR-Sens 3′ UTR vectors with known human miRNA-372, miRNA-373, and miRNA-31 targets (LATS2 and TXNIP). Overall, we observe that miR-Sens- based assays are highly reproducible, allowing detection of the independent contribution of multiple microRNAs to 3′UTR-mediated translational control of LATS2. In conclusion, miR-Sens is a new tool for the efficient study of microRNA activity in primary cells or panels of cell lines. This vector will not only be useful for studies on microRNA biology, but also more broadly on other factors influencing the translation of mRNAs. Copyright © 2012 RNA Society.
dc.description.urihttp://libproxy1.nus.edu.sg/login?url=http://dx.doi.org/10.1261/rna.031831.111
dc.sourceScopus
dc.subjectDual luciferase
dc.subjectMicroRNA
dc.subjectPrimary cells
dc.subjectRetrovirus
dc.subjectUTR
dc.typeArticle
dc.contributor.departmentDUKE-NUS GRADUATE MEDICAL SCHOOL S'PORE
dc.description.doi10.1261/rna.031831.111
dc.description.sourcetitleRNA
dc.description.volume18
dc.description.issue5
dc.description.page1091-1100
dc.description.codenRNARF
dc.identifier.isiut000302909000018
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