title: A non-covalent peptide-based strategy for ex vivo and in vivo oligonucleotide delivery.
abstract: The dramatic acceleration in identification of new nucleic acid-based therapeutic molecules such as short interfering RNA (siRNA) and peptide-nucleic acid (PNA) analogues has provided new perspectives for therapeutic targeting of specific genes responsible for pathological disorders. However, the poor cellular uptake of nucleic acids together with the low permeability of the cell membrane to negatively charged molecules remain major obstacles to their clinical development. Several non-viral strategies have been proposed to improve the delivery of synthetic short oligonucleotides both in cultured cells and in vivo. Cell-penetrating peptides constitute very promising tools for non-invasive cellular import of oligonucleotides and analogs. We recently described a non-covalent strategy based on short amphiphatic peptides (MPG8/PEP3) that have been successfully applied ex vivo and in vivo for the delivery of therapeutic siRNA and PNA molecules. PEP3 and MPG8 form stable nanoparticles with PNA analogues and siRNA, respectively, and promote their efficient cellular uptake, independently of the endosomal pathway, into a wide variety of cell lines, including primary and suspension lines, without any associated cytotoxicity. This chapter describes easy-to-handle protocols for the use of MPG-8 or PEP-3-nanoparticle technologies for PNA and siRNA delivery into adherent and suspension cell lines as well as in vivo into cancer mouse models.
authors: Department of Molecular Biophysics and Therapeutics, Centre de Recherches de Biochimie Macromoléculaire, Montpellier, France. laurence.crombez@crbm.cnrs.fr
authors: Laurence
authors: L
authors: Crombez
authors: May C
authors: MC
authors: Morris
authors: Frederic
authors: F
authors: Heitz
authors: Gilles
authors: G
authors: Divita
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D057846:Cell-Penetrating Peptides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D056744:Cyclin B1 / Q000032:analysis / Q000096:biosynthesis; D004195:Disease Models, Animal; D016503:Drug Delivery Systems / Q000379:methods*; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D053758:Nanoparticles / Q000737:chemistry; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D020135:Peptide Nucleic Acids / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D011092:Polyethylene Glycols / Q000737:chemistry; D011471:Prostatic Neoplasms / Q000188:drug therapy* / Q000473:pathology; D011485:Protein Binding; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism / Q000494:pharmacology*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D014162:Transfection
chemical_list: D057846:Cell-Penetrating Peptides; D056744:Cyclin B1; C543105:MPG-8 peptide; D009841:Oligonucleotides; D009842:Oligopeptides; C520006:Pep-3 peptide; D020135:Peptide Nucleic Acids; D010455:Peptides; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D011092:Polyethylene Glycols