荧光素酶
基因沉默
小RNA
报告基因
生物
基因
分子生物学
非翻译区
转染
基因表达
计算生物学
信使核糖核酸
遗传学
作者
Thomas Clément,Véronique Salone,Mathieu Rederstorff
出处
期刊:Methods in molecular biology
日期:2015-01-01
卷期号:: 187-198
被引量:86
标识
DOI:10.1007/978-1-4939-2547-6_17
摘要
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.
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