Gene Reporter Assays to Study miRNA Function
| Autor: | Yoann Abel, Mathieu Rederstorff |
|---|---|
| Přispěvatelé: | Institut de Génétique Moléculaire de Montpellier (IGMM), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Ingénierie Moléculaire et Physiopathologie Articulaire (IMoPA), Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Untranslated region
Cloning 0303 health sciences Messenger RNA Biology Cell biology 03 medical and health sciences 0302 clinical medicine microRNA Gene silencing Luciferase [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Gene Function (biology) ComputingMilieux_MISCELLANEOUS 030304 developmental biology 030215 immunology |
| Zdroj: | Methods in Molecular Biology; Small non-coding RNAs: Methods and Protocols Methods in Molecular Biology; Small non-coding RNAs: Methods and Protocols, pp.119-131, 2021, ⟨10.1007/978-1-0716-1386-3_12⟩ Methods in Molecular Biology ISBN: 9781071613856 |
| DOI: | 10.1007/978-1-0716-1386-3_12⟩ |
| Popis: | This chapter describes one of the most reliable quantitative assays to test the silencing of a possible target gene by a specific miRNA using a luciferase reporter gene. The experimental procedure first consists in cloning both the wild-type and mutated forms of the 3'UTR of the miRNA-predicted mRNA target downstream of a firefly luciferase reporter. Next, each construct is co-transfected together with the miRNA into HeLa cells, and the reporter expression is monitored. Changes in luciferase levels will indicate whether or not an miRNA can bind to the UTR and regulate its expression. |
| Databáze: | OpenAIRE |
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