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Characterization of Cellular Internalization Pathways for CPP-Mediated Oligonucleotide Delivery

  • Peter Guterstam
  • Samir EL Andaloussi
  • Ülo Langel
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 683)

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.

Key words

Pre-mRNA splicing Oligonucleotide delivery Internalization pathways Endocytosis inhibitors 

Notes

Acknowledgments

pLuc705-HeLa cells were kindly provided by R. Kole and B. Lebleu. This work was supported by the Swedish Science Foundation (VR-NT and VR-MED), the Knut and Alice Wallenberg Foundation, the Swedish Governmental Agency for Innovation Systems (VINNOVA-SAMBIO 2006), and the Swedish Center for Biomembrane Research.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Peter Guterstam
    • 1
  • Samir EL Andaloussi
    • 1
  • Ülo Langel
    • 1
    • 2
  1. 1.Department of NeurochemistryStockholm UniversityStockholmSweden
  2. 2.Laboratory of Molecular Biotechnology, Institute of TechnologyTartu UniversityTartuEstonia

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