Characterization of cellular internalization pathways for CPP-mediated oligonucleotide delivery

Methods Mol Biol. 2011:683:219-30. doi: 10.1007/978-1-60761-919-2_16.

Abstract

The methods for evaluating internalization pathways of cellular CPP-mediated ON delivery utilizing a pre-mRNA splice correction assay and fluorescence-based quantification are described. Examples for characterization of CPP uptake routes, employing various endocytosis inhibitors, and special treatment conditions are demonstrated. The methods are developed to characterize cellular delivery of pre-mRNA splice switching peptide nucleic acids conjugated to CPPs by disulfide bond.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cell-Penetrating Peptides / chemistry
  • Cell-Penetrating Peptides / metabolism*
  • Disulfides / chemistry
  • Drug Carriers / chemistry
  • Drug Carriers / metabolism*
  • Endocytosis / drug effects
  • Fluorometry
  • HeLa Cells
  • Humans
  • Luciferases / genetics
  • Molecular Sequence Data
  • Oligonucleotides / chemistry
  • Oligonucleotides / genetics
  • Oligonucleotides / metabolism*
  • Peptide Nucleic Acids / chemistry
  • Peptide Nucleic Acids / genetics
  • Peptide Nucleic Acids / metabolism
  • Protein Transport
  • RNA Splicing
  • Transfection

Substances

  • Cell-Penetrating Peptides
  • Disulfides
  • Drug Carriers
  • Oligonucleotides
  • Peptide Nucleic Acids
  • Luciferases