Ribonucleoprotein Immunoprecipitation (RNP-IP): A Direct in vivo Analysis of microRNA-Targets

被引：21

作者：

Hassan, Mohammad Q.

Gordon, Jonathan A. R.

Lian, Jane B.

van Wijnen, Andre J.

Stein, Janet L.

Stein, Gary S. ^{[1
]}

机构：

[1] Univ Massachusetts, Sch Med, Dept Cell Biol, Worcester, MA 01655 USA

来源：

JOURNAL OF CELLULAR BIOCHEMISTRY | 2010年 / 110卷 / 04期

关键词：

miR-27A; miR-Let-7/98; RISC; HLX1; RNP-IP; MESSENGER-RNA; IDENTIFICATION; MIRNAS; MECHANISMS; DROSOPHILA; ARGONAUTE2; COMPLEXES;

D O I：

10.1002/jcb.22562

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：

We have developed a novel Ribonucleoprotein Immunoprecipitation (RNP-IP) method to isolate miR-RISC complexes, associated microRNAs and target mRNAs. Our method characterizes mRNAs present in immunoprecipitates containing miR-RISC complexes that were obtained using GW182 and AG02 antibodies. MicroRNA bound transcripts were reverse transcribed and amplified using seed sequence and 3'UTR derived primers. This flexible IP-based assay is a straightforward method to identify miRs participating in gene regulation and their cognate mRNAs in real time. J. Cell. Biochem. 110: 817-822, 2010. (C) 2010 Wiley-Liss, Inc.

引用

收藏

页码：817 / 822

页数：6

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