Direct uptake of Antagomirs and efficient knockdown of miRNA in primary B and T lymphocytes

J Immunol Methods. 2015 Nov:426:128-33. doi: 10.1016/j.jim.2015.07.006. Epub 2015 Jul 15.

Abstract

Depending on their physiological expression level, microRNAs (miRNA) address different target genes, thus have different biological functions. To identify these, the physiological expression has to be blocked. Here, we describe the use of inhibitory cholesterol-modified oligonucleotides (Antagomirs) to inhibit miRNAs selectively in primary human and murine T and B lymphocytes. Due to their lipophilic cholesterol tag Antagomirs enter primary lymphocytes efficiently and quantitatively. We show here that at concentrations of 0.125 to 1μM, Antagomirs selectively inhibit expression of their target miRNA up to 99.5% without affecting cell viability.

Keywords: Antagomir; Murine and human primary lymphocytes; miRNA knockdown.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • B-Lymphocytes / immunology*
  • Biological Transport
  • Cells, Cultured
  • Gene Knockdown Techniques*
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • MicroRNAs / antagonists & inhibitors*
  • MicroRNAs / biosynthesis
  • MicroRNAs / genetics
  • Oligonucleotides / genetics*
  • Spleen / cytology
  • Spleen / immunology
  • Th1 Cells / immunology*

Substances

  • MicroRNAs
  • Oligonucleotides