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Cell Biology & Epigenetics

Products >  Cell_Biology_and_Epigenetics >  RNA_Interference >  MicroRNA >  MirTrap_Overview

Download the MirTrap technical note

The MirTrap System—A powerful tool for discovering and enriching miRNA targets

  • Identification of even transient miRNA targets
  • Efficient pull-down of miRNA targets from all cell types
  • Complete, simplified workflow

What are the challenges in studying microRNA targets?

MicroRNAs are key components in eukaryotic gene expression regulation, guiding the RNA-induced silencing complex (RISC) to a target mRNA. To properly identify and study an miRNA and its target(s), researchers typically rely on immunoprecipitation of the RISC in the hope of also pulling down the bound miRNA/mRNA pair. Herein lies the problem: this interaction is extremely transient (Figure 1), and may not survive the pull-down process. Verification and study of low-abundance miRNA/target mRNAs can be a near impossible task in the face of these circumstances.

Transient interaction of RISC and miRNA/mRNA pair

Figure 1. Transient interaction of RISC and miRNA/mRNA pair, resulting in mRNA processing.

How does the MirTrap System solve the problem?

The MirTrap System (Figure 2) utilizes a dominant negative mutation of a RISC protein subunit (called the “MirTrap protein”). When expressed in mammalian cells, this protein locks your miRNA/mRNA pair into the RISC and limits further processing. A DYKDDDDK (FLAG epitope) tag on the MirTrap protein allows for immunoprecipitation of the complex, which can then be efficiently pulled down without losing your microRNA/target mRNA pairs along the way. Targets that would have been lost using less sensitive methods are instead ready for downstream analysis—either enriching and verifying known miRNA targets with qRT-PCR, or discovering new targets via Next Generation Sequencing (NGS).

Isolation of target mRNAs using the MirTrap System

Figure 2. Experimental workflow for microRNA transfection and isolation of target mRNAs using the MirTrap System.

Read our technical note to get more details and data >>

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