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PepFect6 Mediated SiRNA Delivery into Organotypic Cultures
Stockholm University, Faculty of Science, Department of Neurochemistry.
Number of Authors: 3
2016 (English)In: SiRNA Delivery Methods: Methods and Protocols / [ed] K. Shum, J. Rossi, TOTOWA: HUMANA PRESS INC , 2016, Vol. 1364, 27-35 p.Chapter in book (Refereed)
Abstract [en]

Gene silencing by small interfering RNA (SiRNA) is an attractive therapeutic approach for pathological disorders that targets a specific gene. However, its applications are limited, as naked RNA is rapidly degraded by RNases and is inadequately internalized by the target cells in the body. Several viral and non-viral vectors have been described to improve the delivery of SiRNAs both in cultured cells as well as in vivo. Increasing evidence suggests that cell-penetrating peptides (CPPs) are an efficient, non-cytotoxic tool for intracellular delivery of SiRNA. Recently, a new peptide, PepFect6 (PF6), based system has been described for efficient SiRNA delivery in various cell types. PF6 is an amphipathic stearyl-TP10 peptide carrying a pH titratable trifluoromethylquinoline moiety that facilitate endosomal release. PF6 forms stable non-covalent complexes with SiRNA. Upon internalization, the complexes rapidly escape the endosomal compartment, resulting in robust RNA interference (RNAi) responses. This chapter describes a protocol to use the PF6-nanoparticle technology for SiRNA delivery into organotypic cultures of the inner ear i.e., cochlea. We also highlight different critical points in the peptide/SiRNA complex preparation, transfection and in analyzing the efficacy of PF6-SiRNA associated RNAi response.

Place, publisher, year, edition, pages
TOTOWA: HUMANA PRESS INC , 2016. Vol. 1364, 27-35 p.
Series
Methods in Molecular Biology, ISSN 1064-3745 ; 1364
Keyword [en]
Cell-penetrating peptides (CPPs), PepFect6 (PF6), SiRNA, Inner ear, Cochlea, Organotypic cultures, Fluorescence recovery after photobleaching (FRAP)
National Category
Biological Sciences Environmental Biotechnology
Identifiers
URN: urn:nbn:se:su:diva-128580DOI: 10.1007/978-1-4939-3112-5_3ISI: 000371061200004PubMedID: 26472439ISBN: 978-1-4939-3112-5 (print)ISBN: 978-1-4939-3111-8 (print)OAI: oai:DiVA.org:su-128580DiVA: diva2:916210
Available from: 2016-04-01 Created: 2016-03-30 Last updated: 2016-04-01Bibliographically approved

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