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

Research output: Contribution to journalArticle

19 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)817-822
Number of pages6
JournalJournal of Cellular Biochemistry
Volume110
Issue number4
DOIs
StatePublished - Jul 1 2010
Externally publishedYes

Fingerprint

Ribonucleoproteins
MicroRNAs
Immunoprecipitation
Reduced instruction set computing
Messenger RNA
Gene expression
Seed
Assays
Seeds
Antibodies
Genes

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Hassan, M. Q., Gordon, J. A. R., Lian, J. B., van Wijnen, A. J., Stein, J. L., & Stein, G. S. (2010). Ribonucleoprotein immunoprecipitation (RNP-IP): A direct in vivo analysis of microRNA-targets. Journal of Cellular Biochemistry, 110(4), 817-822. https://doi.org/10.1002/jcb.22562

Ribonucleoprotein immunoprecipitation (RNP-IP) : A direct in vivo analysis of microRNA-targets. / Hassan, Mohammad Q.; Gordon, Jonathan A R; Lian, Jane B.; van Wijnen, Andre J; Stein, Janet L.; Stein, Gary S.

In: Journal of Cellular Biochemistry, Vol. 110, No. 4, 01.07.2010, p. 817-822.

Research output: Contribution to journalArticle

Hassan, MQ, Gordon, JAR, Lian, JB, van Wijnen, AJ, Stein, JL & Stein, GS 2010, 'Ribonucleoprotein immunoprecipitation (RNP-IP): A direct in vivo analysis of microRNA-targets', Journal of Cellular Biochemistry, vol. 110, no. 4, pp. 817-822. https://doi.org/10.1002/jcb.22562
Hassan, Mohammad Q. ; Gordon, Jonathan A R ; Lian, Jane B. ; van Wijnen, Andre J ; Stein, Janet L. ; Stein, Gary S. / Ribonucleoprotein immunoprecipitation (RNP-IP) : A direct in vivo analysis of microRNA-targets. In: Journal of Cellular Biochemistry. 2010 ; Vol. 110, No. 4. pp. 817-822.
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