Improved cell-penetrating peptide–PNA conjugates for splicing redirection in HeLa cells and exon skipping in mdx mouse muscle
| Autor: | Andrey A. Arzumanov, Michael J. Gait, Gabriela D. Ivanova, Matthew J.A. Wood, Bernard Lebleu, Rachida Abes, HaiFang Yin |
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| Přispěvatelé: | Engelhardt Institute of Molecular Biology (ISTC), Russian Academy of Sciences [Moscow] (RAS), Dynamique des interactions membranaires normales et pathologiques (DIMNP), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Montpellier 1 (UM1), HYSED (Hydrodynamics and Sedimentology Laboratory), Department of geography |
| Rok vydání: | 2008 |
| Předmět: |
Peptide Nucleic Acids
mdx mouse Peptide 01 natural sciences Dystrophin Myoblasts Mice chemistry.chemical_compound Exon RNA Precursors MESH: Animals chemistry.chemical_classification MESH: Muscle Skeletal 0303 health sciences Peptide nucleic acid MESH: Peptides MESH: Mice Inbred mdx Exons Transfection MESH: Injections Intramuscular Endocytosis MESH: Endocytosis RNA splicing musculoskeletal diseases RNA Splicing MESH: Biological Transport MESH: RNA Precursors Biology 010402 general chemistry Injections Intramuscular MESH: Peptide Nucleic Acids 03 medical and health sciences MESH: Dystrophin MESH: Muscular Dystrophy Duchenne Genetics Animals Humans [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology RNA Messenger MESH: Myoblasts Muscle Skeletal Molecular Biology MESH: Mice MESH: RNA Messenger 030304 developmental biology MESH: Humans Biological Transport Molecular biology Exon skipping 0104 chemical sciences Muscular Dystrophy Duchenne MESH: Hela Cells chemistry Mice Inbred mdx Cell-penetrating peptide Peptides MESH: RNA Splicing MESH: Exons HeLa Cells |
| Zdroj: | Nucleic Acids Research Nucleic Acids Research, Oxford University Press, 2008, 36 (20), pp.6418-28. ⟨10.1093/nar/gkn671⟩ |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkn671 |
| Popis: | International audience; Steric blocking peptide nucleic acid (PNA) oligonucleotides have been used increasingly for redirecting RNA splicing particularly in therapeutic applications such as Duchenne muscular dystrophy (DMD). Covalent attachment of a cell-penetrating peptide helps to improve cell delivery of PNA. We have used a HeLa pLuc705 cell splicing redirection assay to develop a series of PNA internalization peptides (Pip) conjugated to an 18-mer PNA705 model oligonucleotide with higher activity compared to a PNA705 conjugate with a leading cell-penetrating peptide being developed for therapeutic use, (R-Ahx-R)(4). We show that Pip-PNA705 conjugates are internalized in HeLa cells by an energy-dependent mechanism and that the predominant pathway of cell uptake of biologically active conjugate seems to be via clathrin-dependent endocytosis. In a mouse model of DMD, serum-stabilized Pip2a or Pip2b peptides conjugated to a 20-mer PNA (PNADMD) targeting the exon 23 mutation in the dystrophin gene showed strong exon-skipping activity in differentiated mdx mouse myotubes in culture in the absence of an added transfection agent at concentrations where naked PNADMD was inactive. Injection of Pip2a-PNADMD or Pip2b-PNADMD into the tibealis anterior muscles of mdx mice resulted in approximately 3-fold higher numbers of dystrophin-positive fibres compared to naked PNADMD or (R-Ahx-R)(4)-PNADMD. |
| Databáze: | OpenAIRE |
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