Comparing two approaches of miR-34a target identification, biotinylated-miRNA pulldown vs miRNA overexpression

被引：29

作者：

Awan, Hassaan Mehboob ^{[1
]}

Shah, Abdullah ^{[1
]}

Rashid, Farooq ^{[1
]}

Wei, Shuai ^{[1
]}

Chen, Liang ^{[1
]}

Shan, Ge ^{[1
]}

机构：

[1] Univ Sci & Technol China, Sch Life Sci, CAS Ctr Excellence Mol Cell Sci, CAS Key Lab Innate Immun & Chron Dis, Hefei, Anhui, Peoples R China

来源：

RNA BIOLOGY | 2018年 / 15卷 / 01期

基金：

中国国家自然科学基金;

关键词：

Biotinylated miRNA; Argonaut; 2; microRNA targets; RNA pull down; RNA-seq; MICRORNA TARGETS; CIRCULAR RNAS; ARGONAUTE PROTEINS; TUMOR-SUPPRESSOR; RECOGNITION; EXPRESSION; CELLS; PREDICTION; ROLES;

D O I：

10.1080/15476286.2017.1391441

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：55 / 61

页数：7

共 43 条

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