Silencing of Gene Expression by Gymnotic Delivery of Antisense Oligonucleotides

  • Harris S. Soifer
  • Troels Koch
  • Johnathan Lai
  • Bo Hansen
  • Anja Hoeg
  • Henrik Oerum
  • C. A. Stein
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 815)

Abstract

Antisense oligodeoxyribonucleotides have been used for decades to achieve sequence-specific silencing of gene expression. However, all early generation oligonucleotides (e.g., those with no other modifications than the phosphorothioate backbone) are inactive in vitro unless administered using a delivery vehicle. These delivery vehicles are usually lipidic but can also be polyamines or some other particulate reagent. We have found that by employing locked nucleic acid (LNA) phosphorothioate gap-mer nucleic acids of 16 mer or less in length, and by carefully controlling the plating conditions of the target cells and duration of the experiment, sequence-specific gene silencing can be achieved at low micromolar concentrations in vitro in the absence of any delivery vehicle. This process of naked oligonucleotide delivery to achieve gene silencing in vivo, which we have termed gymnosis, has been observed in many both adherent and nonadherent cell lines against several different targets genes.

Key words

Gymnosis Antisense Silencing Phosphorothioate Locked nucleic acid gap-mer 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Harris S. Soifer
    • 1
  • Troels Koch
    • 2
  • Johnathan Lai
    • 2
  • Bo Hansen
    • 2
  • Anja Hoeg
    • 2
  • Henrik Oerum
    • 2
  • C. A. Stein
    • 3
  1. 1.Department of OncologyMontefi ore Medical Center, Albert Einstein College of MedicineBronxUSA
  2. 2.Santaris PharmaHorsholmDenmark
  3. 3.Albert Einstein College of MedicineAlbert Einstein-Montefi ore Cancer Center, Montefi ore Medical CenterBronxUSA

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