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Related Experiment Videos

Antisense c-myc oligodeoxyribonucleotide cellular uptake.

S Wu-Pong1, T L Weiss, C A Hunt

  • 1Department of Pharmaceutical Chemistry, University of California, San Francisco 94143.

Pharmaceutical Research
|August 1, 1992
PubMed
Summary
This summary is machine-generated.

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Antisense oligonucleotides can inhibit gene expression but how they enter cells is unclear. This study shows oligonucleotide uptake involves surface binding and internalization, possibly via an anion channel, not traditional receptors.

Area of Science:

  • Molecular Biology
  • Genetics

Background:

  • Antisense oligonucleotides (ASOs) show promise for gene expression inhibition.
  • The intracellular delivery mechanism for intact ASOs remains largely unknown.

Purpose of the Study:

  • To investigate the uptake and internalization mechanisms of an oligodeoxyribonucleotide (ODN) targeting the c-myc protooncogene in Rauscher Red 5-1.5 cells.

Main Methods:

  • Utilized a 21-mer ODN complementary to the c-myc translation initiation codon.
  • Assessed surface binding, internalization, and energy dependence of uptake.
  • Investigated the effect of charged molecules, receptor-mediated endocytosis inhibitors, and anion channel inhibitors on uptake.

Main Results:

  • Observed both trypsin-sensitive and trypsin-insensitive surface binding, along with internalization.

Related Experiment Videos

  • Uptake was partially energy-dependent and inhibited by charged molecules (DNA, ATP, dextran sulfate).
  • Traditional receptor-mediated pathways were ruled out; anion channel inhibitors (SITS) and specific salts significantly reduced uptake.

    • Conclusions:

    • Oligonucleotide uptake involves surface binding and internalization, distinct from typical receptor-mediated endocytosis.
    • Evidence suggests a potential role for an anion channel or a novel uptake mechanism.
    • A model was developed to simulate the observed experimental data.