Analysis of microRNA turnover in mammalian cells following Dicer1 ablation

Nucleic Acids Res. 2011 Jul;39(13):5692-703. doi: 10.1093/nar/gkr148. Epub 2011 Mar 29.

Abstract

Although microRNAs (miRNAs) are key regulators of gene expression, little is known of their overall persistence in the cell following processing. Characterization of such persistence is key to the full appreciation of their regulatory roles. Accordingly, we measured miRNA decay rates in mouse embryonic fibroblasts following loss of Dicer1 enzymatic activity. The results confirm the inherent stability of miRNAs, the intracellular levels of which were mostly affected by cell division. Using the decay rates of a panel of six miRNAs representative of the global trend of miRNA decay, we establish a mathematical model of miRNA turnover and determine an average miRNA half-life of 119 h (i.e. ∼5 days). In addition, we demonstrate that select miRNAs turnover more rapidly than others. This study constitutes, to our knowledge, the first in-depth characterization of miRNA decay in mammalian cells. Our findings indicate that miRNAs are up to 10× more stable than messenger RNA and support the existence of novel mechanism(s) controlling selective miRNA cellular concentration and function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • DEAD-box RNA Helicases / genetics*
  • DEAD-box RNA Helicases / metabolism
  • Gene Deletion
  • HEK293 Cells
  • Humans
  • Mice
  • MicroRNAs / metabolism*
  • Models, Biological
  • RNA Processing, Post-Transcriptional
  • RNA Stability*
  • Ribonuclease III / genetics*
  • Ribonuclease III / metabolism

Substances

  • MIRN21 microRNA, mouse
  • MicroRNAs
  • Mirn155 microRNA, mouse
  • Dicer1 protein, mouse
  • Ribonuclease III
  • DEAD-box RNA Helicases