Abstract:
: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 miR-34a.
journal_name
RNA Bioljournal_title
RNA biologyauthors
Awan HM,Shah A,Rashid F,Wei S,Chen L,Shan Gdoi
10.1080/15476286.2017.1391441subject
Has Abstractpub_date
2018-01-02 00:00:00pages
55-61issue
1eissn
1547-6286issn
1555-8584journal_volume
15pub_type
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