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A Non-Covalent Peptide-Based Strategy for siRNA Delivery

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Cell-Penetrating Peptides

Part of the book series: Methods in Molecular Biology ((MIMB,volume 683))

Abstract

The development of short-interfering RNA (siRNA) has provided great hope for therapeutic targeting of specific genes responsible for pathological disorders. However, the poor cellular uptake of siRNA together with the low permeability of the cell membrane to negatively charged molecules, remain major obstacles to clinical development. So far there is no universal method for siRNA delivery as they all present several limitations. Several non-viral strategies have been proposed to improve the delivery of synthetic siRNAs in both cultured cells and in vivo. Cell-penetrating peptides (CPPs) or protein transduction domains (PTD) constitute very promising tools for non-invasive cellular import of siRNA and non-covalent CPP/PTD-based strategies have been successfully applied for ex vivo and in vivo delivery of therapeutic siRNA molecules. We recently described a new peptide-based system, CADY, for efficient delivery of siRNA in both primary and suspension cell lines. CADY is a secondary amphiphatic peptide able to form stable non-covalent complexes with siRNA and to improve their cellular uptake independently of the endosomal pathway. This chapter describes easy to handle protocols for the use of the CADY-nanoparticle technology for the delivery of siRNA into both adherent and suspension cell lines. It will also highlight different critical points in the peptide/siRNA complex preparation and transfection protocols, in order to obtain siRNA-associated interfering response at low nanomolar concentration.

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Acknowledgments

This work was supported in part by the Centre National de la Recherche Scientifique (CNRS), by the Agence Nationale de la Recherche (ANR, ANR-06-BLAN-0071-Pepvec4Ther), and by a grant from Panomics Inc. L.C. was supported by a grant from the Ligue de Recherche contre le Cancer (LNCC). We thank M.C. Morris for critical reading of the manuscript and all members of the laboratory for fruitful discussions.

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Authors and Affiliations

  1. Department of Molecular Biophysics and Therapeutics, Centre de Recherches de Biochimie Macromoléculaire, Montpellier, France

    Laurence Crombez

  2. Biophysics Department, Centre de Recherches de Biochimie Macromoléculaire, Montpellier University, Montpellier, France

    Gilles Divita

Authors
  1. Laurence Crombez
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  2. Gilles Divita
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Correspondence to Gilles Divita .

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Editors and Affiliations

  1. , Department of Neurochemistry, Stockholm University, Svante Arrheniusv 21A21A, Stockholm, 106 91, Sweden

    Ülo Langel

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Crombez, L., Divita, G. (2011). A Non-Covalent Peptide-Based Strategy for siRNA Delivery. In: Langel, Ü. (eds) Cell-Penetrating Peptides. Methods in Molecular Biology, vol 683. Humana Press. https://doi.org/10.1007/978-1-60761-919-2_25

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  • DOI: https://doi.org/10.1007/978-1-60761-919-2_25

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-918-5

  • Online ISBN: 978-1-60761-919-2

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