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

页码：817 / 822

页数：6

共 12 条

[1] Microarray identification of FMRP-associated brain mRNAs and altered mRNA translational profiles in fragile X syndrome
Brown, V
Jin, P
Ceman, S
Darnell, JC
O'Donnell, WT
Tenenbaum, SA
Jin, XK
Feng, Y
Wilkinson, KD
Keene, JD
Darnell, RB
Warren, ST
[J]. CELL, 2001, 107 (04) : 477 - 487
[2] Extensive association of functionally and cytotopically related mRNAs with Puf family RNA-binding proteins in yeast
Gerber, AP
Herschlag, D
Brown, PO
[J]. PLOS BIOLOGY, 2004, 2 (03): : 342 - 354
[3] Genome-wide identification of mRNAs associated with the translational regulator PUMILIO in Drosophila melanogaster
Gerber, AP
Luschnig, S
Krasnow, MA
Brown, PO
Herschlag, D
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (12) : 4487 - 4492
[4] RIP-Chip: the isolation and identification of mRNAs, microRNAs and protein components of ribonucleoprotein complexes from cell extracts
Keene, Jack D.
Komisarow, Jordan M.
Friedersdorf, Matthew B.
[J]. NATURE PROTOCOLS, 2006, 1 (01) : 302 - 307
[5] Biological Functions of miR-29b Contribute to Positive Regulation of Osteoblast Differentiation
Li, Zhaoyong
Hassan, Mohammad Q.
Jafferji, Mohammed
Aqeilan, Rami I.
Garzon, Ramiro
Croce, Carlo M.
van Wijnen, Andre J.
Stein, Janet L.
Stein, Gary S.
Lian, Jane B.
[J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2009, 284 (23) : 15676 - 15684
[6] Argonaute2 is the catalytic engine of mammalian RNAi
Liu, JD
Carmell, MA
Rivas, FV
Marsden, CG
Thomson, JM
Song, JJ
Hammond, SM
Joshua-Tor, L
Hannon, GJ
[J]. SCIENCE, 2004, 305 (5689) : 1437 - 1441
[7] Meister G, 2004, MOL CELL, V15, P185, DOI 10.1016/j.molcel.2004.07.007
[8] Repression of protein synthesis by miRNAs: how many mechanisms?
Pillai, Ramesh S.
Bhattacharyya, Suvendra N.
Filipowicz, Witold
[J]. TRENDS IN CELL BIOLOGY, 2007, 17 (03) : 118 - 126
[9] MicroRNA function: Multiple mechanisms for a tiny RNA?
Pillai, RS
[J]. RNA, 2005, 11 (12) : 1753 - 1761
[10] Identifying mRNA subsets in messenger ribonucleoprotein complexes by using cDNA arrays
Tenenbaum, SA
Carson, CC
Lager, PJ
Keene, JD
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (26) : 14085 - 14090

← 1 2 →