title: Optimisation of vectorisation property: A comparative study for a secondary amphipathic peptide.
abstract: RNA interference provides a powerful technology for specific gene silencing. Therapeutic applications of small interfering RNA (siRNA) however require efficient vehicles for stable complexation and intracellular delivery. In order to enhance their cell delivery, short amphipathic peptides called cell-penetrating peptides (CPPs) have been intensively developed for the last two decades. In this context, the secondary amphipathic peptide CADY has shown to form stable siRNA complexes and to improve their cellular uptake independent of the endosomal pathway. In the present work, we have described the parameters influencing CADY nanoparticle formation (buffers, excipients, presence of serum, etc.), and have followed in details the CPP:siRNA self-assembly. Once optimal conditions were determined, we have compared the ability of seven different CADY analogues to form siRNA-loaded nanoparticles compared to CADY:siRNA. First of all, we were able to show by biophysical methods that structural polymorphism (α-helix) is an important prerequisite for stable nanoparticle formation independently of occurring sequence mutations. Luciferase assays revealed that siRNA complexed to CADY-K (shorter version) shows better knock-down efficiency on Neuro2a-Luc(+) and B16-F10-Luc(+) cells compared to CADY:siRNA. Altogether, CADY-K is an ideal candidate for further application especially with regards to ex vivo or in vivo applications.
authors.affiliation: Centre de Recherche de Biologie cellulaire de Montpellier (CRBM), CNRS UMR 5237, 1919 Route de Mende, 34293 Montpellier CEDEX 5, France.
authors.forename: Karidia
authors.initials: K
authors.lastname: Konate
authors.affiliation: Centre de Recherche de Biologie cellulaire de Montpellier (CRBM), CNRS UMR 5237, 1919 Route de Mende, 34293 Montpellier CEDEX 5, France.
authors.forename: Mattias F
authors.initials: MF
authors.lastname: Lindberg
authors.affiliation: Centre de Recherche de Biologie cellulaire de Montpellier (CRBM), CNRS UMR 5237, 1919 Route de Mende, 34293 Montpellier CEDEX 5, France.
authors.forename: Anaïs
authors.initials: A
authors.lastname: Vaissiere
authors.affiliation: Centre de Recherche de Biologie cellulaire de Montpellier (CRBM), CNRS UMR 5237, 1919 Route de Mende, 34293 Montpellier CEDEX 5, France.
authors.forename: Carole
authors.initials: C
authors.lastname: Jourdan
authors.affiliation: Sys2Diag, FRE3690-CNRS/ALCEDIAG, 1682 Rue de la Valsiere, 34184 Montpellier CEDEX 4, France.
authors.forename: Gudrun
authors.initials: G
authors.lastname: Aldrian
authors.affiliation: CNRS UMR5048, Centre de Biochimie Structurale, 29 rue de Navacelles, 34090 Montpellier, France; INSERM U1054, 34090 Montpellier, France; Université de Montpellier, 34090 Montpellier, France.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Margeat
authors.affiliation: Centre de Recherche de Biologie cellulaire de Montpellier (CRBM), CNRS UMR 5237, 1919 Route de Mende, 34293 Montpellier CEDEX 5, France. Electronic address: sebastien.deshayes@crbm.cnrs.fr.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Deshayes
authors.affiliation: Centre de Recherche de Biologie cellulaire de Montpellier (CRBM), CNRS UMR 5237, 1919 Route de Mende, 34293 Montpellier CEDEX 5, France. Electronic address: prisca.boisguerin@crbm.cnrs.fr.
authors.forename: Prisca
authors.initials: P
authors.lastname: Boisguerin
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D057846:Cell-Penetrating Peptides / Q000737:chemistry*; D004337:Drug Carriers / Q000737:chemistry; D020868:Gene Silencing / Q000187:drug effects; D057927:Hydrophobic and Hydrophilic Interactions; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry; D034622:RNA Interference / Q000502:physiology; D034741:RNA, Small Interfering / Q000378:metabolism
chemical_list: D057846:Cell-Penetrating Peptides; D004337:Drug Carriers; D034741:RNA, Small Interfering
keywords: Cell penetrating peptide; Circular dichroism; Dynamic light scattering; Fluorescence correlation spectroscopy; Luciferase assay; Nanoparticle; Self-assembly; siRNA