Dual luciferase gene reporter assays to study miRNA function

Methods in Molecular Biology
Thomas ClémentMathieu Rederstorff

Abstract

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 a miRNA can bind to the UTR and regulate its expression.

Citations

Mar 29, 2019·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Arie FridrichYehu Moran
Dec 25, 2019·Respirology : Official Journal of the Asian Pacific Society of Respirology·Yasuhiro GonMari Hikichi
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Jun 24, 2020·Biological Trace Element Research·Chen LiXueli Pan
Mar 16, 2017·Proceedings of the National Academy of Sciences of the United States of America·Xiaodan Wang, Dietmar Kültz
Feb 6, 2020·Clinical and Translational Medicine·Estanislao NavarroMiguel Hueso
Jan 12, 2021·Frontiers in Cell and Developmental Biology·Fei YanYuan Wu
Nov 19, 2020·International Journal of Molecular Sciences·Adrian Pérez-AgustínSara Pagans
Feb 5, 2021·Frontiers in Bioengineering and Biotechnology·Tarana Siddika, Ilka U Heinemann
Dec 12, 2020·Nucleic Acids Research·Fabian KernEckart Meese
Mar 9, 2021·Computational and Structural Biotechnology Journal·Petr V Nazarov, Stephanie Kreis

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