Abstract
Amphipathic, pH-responsive, membrane-active peptides such as LAH4 and derivatives thereof have the ability to effectively deliver genes and small interfering RNA (siRNA) into mammalian cells. Their ability to bind and protect nucleic acids and then disrupt membranes when activated at low pH enables them to harness the endocytic machinery to deliver cargo efficiently and with low associated toxicity. This chapter describes protocols for the chemical synthesis of transfection peptides of the LAH4 family, complex formation with nucleic acids, and their use for the in vitro delivery of either plasmid DNA or siRNA into mammalian cell lines.
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Acknowledgments
We thank Christopher Aisenbrey for his assistance for the section concerning the chemical synthesis of the peptides. We are most grateful for the financial support by the Agence Nationale de la Recherche (TRANSPEP, project ANR-07-PCVI-0018), Vaincre la Mucoviscidose, the University of Strasbourg (PPF RMN), the Region Alsace, the French Ministry of Research, the CNRS, and the Medical Research Council (New Investigator Research Grant to AJM).
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Kichler, A., Mason, A.J., Marquette, A., Bechinger, B. (2013). Histidine-Rich Cationic Amphipathic Peptides for Plasmid DNA and siRNA Delivery. In: Ogris, M., Oupicky, D. (eds) Nanotechnology for Nucleic Acid Delivery. Methods in Molecular Biology, vol 948. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-140-0_7
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DOI: https://doi.org/10.1007/978-1-62703-140-0_7
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-139-4
Online ISBN: 978-1-62703-140-0
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