title: Comparison of novel delivery systems for antisense peptide nucleic acids.
abstract: Peptide nucleic acids (PNAs) provide a powerful tool to study the mechanism of transcription and translation, an innovative strategy to regulate target gene expression. They have been successfully used in antisense technology, for their ability to specifically bind to messenger RNA (mRNA) targets and to inhibit translation of the target genes. However, unlike most of the DNA and RNA oligonucleotides, PNAs are poorly penetrated through the cell membrane, partially due to their uncharged property. To enhance the efficiency in PNA delivery, many strategies have been explored. We here compare the efficacy of three different delivery strategies for antisense PNA: 1) conjugation to hydrophobic peptides, 2) adsorption onto polymeric microspheres and 3) encapsulation in autologous erythrocytes. To this purpose, we designed and prepared PNA sequences able to inhibit the expression of macrophage enzymes involved in inflammatory process, i.e. nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) and tested their antisense activity in a murine macrophage cellular model. Both delivery through polymeric microspheres and encapsulation into erythrocytes allowed the antisense activity of unmodified PNAs at nanomolar concentration.
authors.affiliation: Institute of Biochemistry Giorgio Fornaini, University of Urbino Carlo Bo, Italy. l.chiarantini@uniurb.it
authors.forename: Laura
authors.initials: L
authors.lastname: Chiarantini
authors.forename: Aurora
authors.initials: A
authors.lastname: Cerasi
authors.forename: Alessandra
authors.initials: A
authors.lastname: Fraternale
authors.forename: Enrico
authors.initials: E
authors.lastname: Millo
authors.forename: Umberto
authors.initials: U
authors.lastname: Benatti
authors.forename: Katia
authors.initials: K
authors.lastname: Sparnacci
authors.forename: Michele
authors.initials: M
authors.lastname: Laus
authors.forename: Marco
authors.initials: M
authors.lastname: Ballestri
authors.forename: Luisa
authors.initials: L
authors.lastname: Tondelli
mesh_terms: D000818:Animals; D016375:Antisense Elements (Genetics); D055598:Chemical Phenomena; D002627:Chemistry, Physical; D051546:Cyclooxygenase 2 / Q000096:biosynthesis / Q000235:genetics; D004339:Drug Compounding; D016503:Drug Delivery Systems; D004912:Erythrocytes / Q000201:enzymology / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D066298:In Vitro Techniques; D007249:Inflammation / Q000235:genetics / Q000473:pathology; D008262:Macrophage Activation; D008264:Macrophages / Q000201:enzymology; D051379:Mice; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D052247:Nitric Oxide Synthase Type II / Q000096:biosynthesis / Q000235:genetics; D009573:Nitrites / Q000378:metabolism; D020135:Peptide Nucleic Acids / Q000008:administration & dosage* / Q000235:genetics; D010587:Phagocytosis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D016375:Antisense Elements (Genetics); D009573:Nitrites; D020135:Peptide Nucleic Acids; D052247:Nitric Oxide Synthase Type II; D051546:Cyclooxygenase 2