BMC Research Notes | |
Validation of miRNA-mRNA interactions by electrophoretic mobility shift assays | |
Carlos J Ciudad1  Véronique Noé1  Xenia Villalobos1  Núria Mencia1  Anna Solé1  | |
[1] Department of Biochemistry and Molecular Biology, School of Pharmacy, University of Barcelona, Avenue Diagonal 643, Barcelona E-08028, Spain | |
关键词: EMSA; Target validation; miRNA; Binding assay; 3′-UTR; | |
Others : 1140872 DOI : 10.1186/1756-0500-6-454 |
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received in 2013-06-20, accepted in 2013-11-08, 发布年份 2013 | |
【 摘 要 】
Background
MicroRNAs are small non-coding RNAs involved in gene expression regulation by targeting specific regions in the 3′-UTR of the mRNA of their target genes. This binding leads to a decrease in the protein levels of such genes either by mRNA degradation or mRNA destabilization and translation inhibition. The interaction between a miRNA and its target mRNAs is usually studied by co-transfection of a reporter expression vector containing the 3′-UTR region of the mRNA and an inhibitory or precursor molecule for the miRNA. This approach, however, does not measure the direct and physical interaction between a miRNA and a specific mRNA.
Findings
RNA molecules corresponding to miR-224 and to the 3′-UTR of SLC4A4 were incubated together and their interaction studied under different binding conditions using electrophoretic mobility shift assays. A direct and specific interaction between miR-224 and SLC4A4 mRNA was observed. This interaction was abolished in the presence of competitors.
Conclusions
In this study, we explored a new application for the electrophoretic mobility shift assay and we demonstrated that it is a useful alternative method to assess, in a direct and specific manner, whether a miRNA binds to a specific predicted target mRNA.
【 授权许可】
2013 Solé et al.; licensee BioMed Central Ltd.
【 预 览 】
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20150325133944529.pdf | 1680KB | download | |
Figure 4. | 83KB | Image | download |
Figure 3. | 43KB | Image | download |
Figure 2. | 75KB | Image | download |
Figure 1. | 65KB | Image | download |
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