4.2 Article

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

Journal

JOURNAL OF IMMUNOLOGICAL METHODS
Volume 426, Issue -, Pages 128-133

Publisher

ELSEVIER SCIENCE BV
DOI: 10.1016/j.jim.2015.07.006

Keywords

miRNA knockdown; Antagomir; Murine and human primary lymphocytes

Funding

  1. Deutsche Forschungsgemeinschaft [SFB 650, SFB 633]
  2. IMI JU funded project BTCure [115241]
  3. FORSYS (Forschungseinheiten zur Systembiologie) program
  4. Federal Ministry of Education and Research
  5. European Research Council (ERC) Advanced Grant [ERC-2010-AdG_20100317, 268987]
  6. European Research Council (ERC) [268987] Funding Source: European Research Council (ERC)

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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 1 mu M, Antagomirs selectively inhibit expression of their target miRNA up to 99.5% without affecting cell viability. (C) 2015 The Authors. Published by Elsevier B.V.

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