caa a22 4500 445326646 CHVBK 20180317142723.0 cr unu---uuuuu 170323e20110601xx s 000 0 eng 10.1007/s10989-011-9252-1 doi (NATIONALLICENCE)springer-10.1007/s10989-011-9252-1 NickFects, Phosphorylated Derivatives of Transportan 10 for Cellular Delivery of Oligonucleotides [Elektronische Daten] [Nikita Oskolkov, Piret Arukuusk, Dana-Maria Copolovici, Staffan Lindberg, Helerin Margus, Kärt Padari, Margus Pooga, Ülo Langel] Oligonucleotide-based gene regulation has a high potential in gene therapy, but the plasma membrane is impermeable for nucleic acid polymers and, consequently, an efficient and non-toxic transfection agent is needed for their delivery into the cell. In this study we present a novel series, NickFects, of chemically modified TP10 peptide-based delivery vectors used for the cellular delivery of single-stranded oligonucleotides. These carriers, obtained by replacement of Ile8 by threonine in stearyl-TP10 and by modifying of tyrosine and/or threonine, respectively, by phosphorylation formed 300-500nm in size peptide:oligonucleotide nanocomplexes with negative surface charges. The highest splice-correcting effect was obtained when phosphorotiate 2′-O-methyl oligonucleotides were transduced into cells by NickFect1 (NF1) or NickFect2 (NF2). In addition, we also show how a small modification (one or two negative charges) in peptide sequence can affect its ability to deliver ONs into cells and increase their potency in the splicing redirection assay. Our studies demonstrate that NF1 and NF2 have higher transfection efficacy for oligonucleotides as compared to the most commonly used transfection agent Lipofectamine™ 2000 and lead to higher biological response in cells. Springer Science+Business Media, LLC, 2011 Cell-penetrating peptide nationallicence Splice correction assay nationallicence Co-incubation nationallicence Stearylation nationallicence Phosphorylated peptide nationallicence Delivery vector nationallicence 2′-OMe ON : Phosphorothioate 2′-O-methyl RNA nationallicence SCO : Splice-correcting oligonucleotides nationallicence CPPs : Cell-penetrating peptides nationallicence PBS : Phosphate buffered saline nationallicence TFA : Trifluoroacetic acid nationallicence TP10 : Transportan 10 nationallicence ON : Oligonucleotide nationallicence SFM : Serum-free medium nationallicence FM : Serum-containing medium nationallicence OD : Optical density nationallicence HOBt : Hydroxybenzotriazole nationallicence HBTU : O-Benzotriazole-N,N,N′,N′-tetramethyl-uronium-hexafluoro-phosphate nationallicence DIEA : Diisopropylethylamine nationallicence TIS : Triisopropylsilane nationallicence Oskolkov Nikita Laboratory of Molecular Biotechnology, Institute of Technology, University of Tartu, Nooruse 1, 50411, Tartu, Estonia aut Arukuusk Piret Laboratory of Molecular Biotechnology, Institute of Technology, University of Tartu, Nooruse 1, 50411, Tartu, Estonia aut Copolovici Dana-Maria Laboratory of Molecular Biotechnology, Institute of Technology, University of Tartu, Nooruse 1, 50411, Tartu, Estonia aut Lindberg Staffan Department of Neurochemistry, The Arrhenius Laboratories for Natural Sciences, Stockholm University, Svante Arrheniusväg 21A, 10691, Stockholm, Sweden aut Margus Helerin Department of Developmental Biology, Institute of Molecular and Cell Biology, University of Tartu, 51010, Tartu, Estonia aut Padari Kärt Department of Developmental Biology, Institute of Molecular and Cell Biology, University of Tartu, 51010, Tartu, Estonia aut Pooga Margus Department of Developmental Biology, Institute of Molecular and Cell Biology, University of Tartu, 51010, Tartu, Estonia aut Langel Ülo Laboratory of Molecular Biotechnology, Institute of Technology, University of Tartu, Nooruse 1, 50411, Tartu, Estonia aut International Journal of Peptide Research and Therapeutics Springer Netherlands 17/2(2011-06-01), 147-157 1573-3149 17:2<147 2011 17 10989 https://doi.org/10.1007/s10989-011-9252-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10989-011-9252-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Oskolkov Nikita Laboratory of Molecular Biotechnology, Institute of Technology, University of Tartu, Nooruse 1, 50411, Tartu, Estonia aut NATIONALLICENCE 700 1- Arukuusk Piret Laboratory of Molecular Biotechnology, Institute of Technology, University of Tartu, Nooruse 1, 50411, Tartu, Estonia aut NATIONALLICENCE 700 1- Copolovici Dana-Maria Laboratory of Molecular Biotechnology, Institute of Technology, University of Tartu, Nooruse 1, 50411, Tartu, Estonia aut NATIONALLICENCE 700 1- Lindberg Staffan Department of Neurochemistry, The Arrhenius Laboratories for Natural Sciences, Stockholm University, Svante Arrheniusväg 21A, 10691, Stockholm, Sweden aut NATIONALLICENCE 700 1- Margus Helerin Department of Developmental Biology, Institute of Molecular and Cell Biology, University of Tartu, 51010, Tartu, Estonia aut NATIONALLICENCE 700 1- Padari Kärt Department of Developmental Biology, Institute of Molecular and Cell Biology, University of Tartu, 51010, Tartu, Estonia aut NATIONALLICENCE 700 1- Pooga Margus Department of Developmental Biology, Institute of Molecular and Cell Biology, University of Tartu, 51010, Tartu, Estonia aut NATIONALLICENCE 700 1- Langel Ülo Laboratory of Molecular Biotechnology, Institute of Technology, University of Tartu, Nooruse 1, 50411, Tartu, Estonia aut NATIONALLICENCE 773 0- International Journal of Peptide Research and Therapeutics Springer Netherlands 17/2(2011-06-01), 147-157 1573-3149 17:2<147 2011 17 10989 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer