title: A stearylated CPP for delivery of splice correcting oligonucleotides using a non-covalent co-incubation strategy.
abstract: Aberrations in splicing patterns play a significant role in several diseases, and splice correction, together with other forms of gene regulation, is consequently an emerging therapeutic target. In order to achieve successful oligonucleotide transfection, efficient delivery vectors are generally necessary. In this study we present one such vector, the chemically modified cell-penetrating peptide (CPP) TP10, for efficient delivery of a splice-correcting 2'-OMe RNA oligonucleotide. Utilizing a functional splice correction assay, we assessed the transfection efficiency of non-covalent complexes of oligonucleotides and stearylated or cysteamidated CPPs. Stearylation of the CPPs Arg9 and penetratin, as well as cysteamidation of MPG and TP10, did not improve transfection, whereas the presence of an N-terminal stearyl group on TP10 improved delivery efficiency remarkably compared to the unmodified peptide. The splice correction levels observed with stearyl-TP10 are in fact in parity with the effects seen with the commercially available transfection agent Lipofectamine 2000. However, the inherent toxicity associated with cationic lipid-based transfections can be completely eliminated when using the stearylated TP10, making this vector highly promising for non-covalent delivery of negatively charged oligonucleotides.
authors.affiliation: Department of Neurochemistry, the Arrhenius Laboratories for Natural Sciences, Stockholm University, Svante Arrheniuväg 21A, SE-10691 Stockholm, Sweden. maarja@neurochem.su.se
authors.forename: Maarja
authors.initials: M
authors.lastname: Mäe
authors.forename: Samir
authors.initials: S
authors.lastname: El Andaloussi
authors.forename: Per
authors.initials: P
authors.lastname: Lundin
authors.forename: Nikita
authors.initials: N
authors.lastname: Oskolkov
authors.forename: Henrik J
authors.initials: HJ
authors.lastname: Johansson
authors.forename: Peter
authors.initials: P
authors.lastname: Guterstam
authors.forename: Ulo
authors.initials: U
authors.lastname: Langel
mesh_terms: D017398:Alternative Splicing / Q000187:drug effects* / Q000235:genetics; D018929:Cell Culture Techniques; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002738:Chloroquine / Q000494:pharmacology; D003470:Culture Media; D019004:Galanin / Q000737:chemistry*; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D008055:Lipids / Q000737:chemistry; D054735:Phosphorothioate Oligonucleotides / Q000008:administration & dosage* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D013229:Stearic Acids / Q000737:chemistry*; D014162:Transfection; D014862:Wasp Venoms / Q000737:chemistry*
chemical_list: D003470:Culture Media; D008055:Lipids; C086724:Lipofectamine; D054735:Phosphorothioate Oligonucleotides; D011993:Recombinant Fusion Proteins; D013229:Stearic Acids; D014862:Wasp Venoms; C110068:transportan; C031183:stearic acid; D002738:Chloroquine; D019004:Galanin