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The Assembly of Fluorescently Labeled Peptide–Oligonucleotide Conjugates via Orthogonal Ligation Strategies

  • John Karas
  • Bradley J. Turner
  • Fazel ShabanpoorEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1828)

Abstract

Efficient intracellular delivery is critical to the successful application of synthetic antisense oligonucleotides (ASOs) to modulate gene expression. The conjugation of cell-penetrating peptides (CPPs) to ASOs has been shown to significantly improve their intracellular delivery. It is important, however, that formation of the covalent linkage between the peptide and oligonucleotide is efficient and orthogonal, to ensure high yields and a homogeneous product. Described herein are efficient and facile methodologies for the conjugation of peptides to ASOs, and their subsequent labeling with various moieties such as fluorescent dyes for intracellular tracking studies.

Key words

Peptides Oligonucleotides Conjugation Fluorescent labeling Alkyne–azide 

Notes

Acknowledgment

This work was supported by MND Research Institute of Australia (MNDRIA GIA-1722), National Health and Medical Research Council (Project Grant 1104299), and the Stafford Fox Medical Research Foundation.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • John Karas
    • 1
  • Bradley J. Turner
    • 2
  • Fazel Shabanpoor
    • 2
    • 3
    Email author
  1. 1.Pharmacology and TherapeuticsThe University of MelbourneMelbourneAustralia
  2. 2.The Florey Institute of Neuroscience and Mental HealthThe University of MelbourneMelbourneAustralia
  3. 3.School of ChemistryThe University of MelbourneMelbourneAustralia

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