A biochemical approach to identify direct microRNA targets

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

Research output: Contribution to journalArticle

15 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
Pages (from-to)29-37
Number of pages9
JournalMethods in Molecular Biology
Volume1206
DOIs
Publication statusPublished - 1 Jan 2015
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

  • Molecular Biology
  • Genetics

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