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Non-covalent Encapsulation of siRNA with Cell-Penetrating Peptides

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Design and Delivery of SiRNA Therapeutics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2282))

Abstract

SiRNAs may act as selective and potent therapeutics, but poor deliverability in vivo is a limitation. Among the recently proposed vectors, cell-penetrating peptides (CPPs), also referred as protein transduction domains (PTDs), allow siRNA stabilization and increased cellular uptake. This chapter aims to guide scientists in the preparation and characterization of CPP-siRNA complexes, particularly the evaluation of novel CPPs variants for siRNA encapsulation and delivery. Herein, we present a collection of methods to determine CPP-siRNA interaction, encapsulation, stability, conformation, transfection, and silencing efficiency.

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Acknowledgements

We gratefully acknowledge funding support from the European Union PathChooser Marie-Curie Initial Training Network (ITN)(PITN-GA-2013-608373), the Lundbeckfonden Center of Excellence NanoCAN, the University of Southern Denmark (SDU) DAWN-2020 project of the SDU Presidents SDU2020 program, the Danish Cancer Society, A Race Agaisnt Breast Cancer and the A.P. Moeller Foundation. Moreover, we acknowledge the Danish Molecular Biomedical Imaging Center (DaMBIC; University of Southern Denmark) for the use of the bioimaging facilities, and Prof. Jesper Wengel (Biomolecular Nanoscale Engineering Center, University of Southern Denmark) for providing the tdTomato1 siRNA.

Conflicts of Interest: The authors declare no conflict of interest.

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Authors and Affiliations

  1. Department of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark

    Martina Tuttolomondo & Henrik J. Ditzel

  2. Department of Oncology, Odense University Hospital, Odense, Denmark

    Henrik J. Ditzel

Authors
  1. Martina Tuttolomondo
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  2. Henrik J. Ditzel
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Corresponding authors

Correspondence to Martina Tuttolomondo or Henrik J. Ditzel .

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Editors and Affiliations

  1. Department of Oncology, Odense University Hospital Odense, Denmark, Department of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark

    Henrik J. Ditzel

  2. Department of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark

    Martina Tuttolomondo

  3. Center for RNA Medicine, Department of Clinical Medicine, Aalborg University, Copenhagen, Denmark

    Sakari Kauppinen

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Tuttolomondo, M., Ditzel, H.J. (2021). Non-covalent Encapsulation of siRNA with Cell-Penetrating Peptides. In: Ditzel, H.J., Tuttolomondo, M., Kauppinen, S. (eds) Design and Delivery of SiRNA Therapeutics. Methods in Molecular Biology, vol 2282. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1298-9_19

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  • DOI: https://doi.org/10.1007/978-1-0716-1298-9_19

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1297-2

  • Online ISBN: 978-1-0716-1298-9

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