A biochemical approach to identify direct microRNA targets

Murugan Subramanian, Xiao Ling Li, Toshifumi Hara, Ashish Lal

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Citations (Scopus)

Abstract

We have recently developed a biochemical approach to isolate miRNA-bound mRNAs and have used this method to identify the genome-wide mRNAs regulated by the tumor suppressor miRNA miR-34a. This method involves transfection of cells with biotinylated miRNA mimics, streptavidin pulldown, RNA isolation, and qRT-PCR. The protocol in this chapter describes these steps and the issues that should be considered while designing such pulldown experiments.

Original languageEnglish
Title of host publicationRegulatory Non-Coding RNAs
Subtitle of host publicationMethods and Protocols
PublisherSpringer New York
Pages29-37
Number of pages9
ISBN (Electronic)9781493913695
ISBN (Print)9781493913688
DOIs
Publication statusPublished - 20 Sep 2014
Externally publishedYes

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Keywords

  • Argonaute
  • Dicer
  • MicroRNA
  • MicroRNP
  • Native agarose gel
  • Native PAGE
  • Pre-miRNA
  • Ribonucleoprotein
  • RISC
  • RNA-protein

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Subramanian, M., Li, X. L., Hara, T., & Lal, A. (2014). A biochemical approach to identify direct microRNA targets. In Regulatory Non-Coding RNAs: Methods and Protocols (pp. 29-37). Springer New York. https://doi.org/10.1007/978-1-4939-1369-5_3