期刊
RNA BIOLOGY
卷 15, 期 1, 页码 55-61出版社
TAYLOR & FRANCIS INC
DOI: 10.1080/15476286.2017.1391441
关键词
Biotinylated miRNA; Argonaut 2; microRNA targets; RNA pull down; RNA-seq
资金
- National Basic Research Program of China [2015CB943000]
- National Natural Science Foundation of China [31471225, 31600657]
- Major / Innovative Program of Development Foundation of Hefei Center for Physical Science and Technology [2016FXCX006]
- Open Project of the CAS Key Laboratory of Innate Immunity and Chronic Disease [KLIICD-201603]
microRNAs (miRNAs) are critical regulators of gene expression. For elucidating functional roles of miRNAs, it is critical to identify their direct targets. There are debates about whether pulldown of biotinylated miRNA mimics can be used to identify miRNA targets or not. Here we show that biotin-labelled miR-34a can be loaded to AGO2, and AGO2 immunoprecipitation can pulldown biotinylated miR-34a (Bio-miR pulldown). RNA-sequencing (RNA-seq) of the Bio-miR pulldown RNAs efficiently identified miR-34a mRNA targets, which could be verified with luciferase assays. In contrast to the approach of Bio-miR pulldown, RNA-seq of miR-34a overexpression samples had limited value in identifying direct targets of miR-34a. It seems that pulldown of 3'-Biotin-tagged miRNA can identify bona fide microRNA targets at least for miR34a.
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