Ribonucleoprotein immunoprecipitation (RNP-IP): A direct in vivo analysis of microRNA-targets

Mohammad Q. Hassan; Jonathan A.R. Gordon; Jane B. Lian; Andre J. Van Wijnen; Janet L. Stein; Gary S. Stein

doi:10.1002/jcb.22562

Ribonucleoprotein immunoprecipitation (RNP-IP): A direct in vivo analysis of microRNA-targets

Mohammad Q. Hassan, Jonathan A.R. Gordon, Jane B. Lian, Andre J. Van Wijnen, Janet L. Stein, Gary S. Stein

Orthopedic Surgery

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21 Scopus citations

Abstract

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 AGO2 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.

Original language	English (US)
Pages (from-to)	817-822
Number of pages	6
Journal	Journal of cellular biochemistry
Volume	110
Issue number	4
DOIs	https://doi.org/10.1002/jcb.22562
State	Published - Jul 1 2010

Keywords

HLX1
RISC
RNP-IP
miR-27A
miR-Let-7/98

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Cell Biology

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10.1002/jcb.22562

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title = "Ribonucleoprotein immunoprecipitation (RNP-IP): A direct in vivo analysis of microRNA-targets",

abstract = "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 AGO2 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.",

keywords = "HLX1, RISC, RNP-IP, miR-27A, miR-Let-7/98",

author = "Hassan, {Mohammad Q.} and Gordon, {Jonathan A.R.} and Lian, {Jane B.} and {Van Wijnen}, {Andre J.} and Stein, {Janet L.} and Stein, {Gary S.}",

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T2 - A direct in vivo analysis of microRNA-targets

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AU - Gordon, Jonathan A.R.

AU - Lian, Jane B.

AU - Van Wijnen, Andre J.

AU - Stein, Janet L.

AU - Stein, Gary S.

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AB - 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 AGO2 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.

KW - HLX1

KW - RISC

KW - RNP-IP

KW - miR-27A

KW - miR-Let-7/98

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JF - Journal of cellular biochemistry

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Ribonucleoprotein immunoprecipitation (RNP-IP): A direct in vivo analysis of microRNA-targets

Abstract

Keywords

ASJC Scopus subject areas

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