title: Evaluation of transportan 10 in PEI mediated plasmid delivery assay.
abstract: Cell-penetrating peptides (CPPs) are novel high-capacity delivery vectors for different bioactive cargoes. We have evaluated the CPP transportan 10 (TP10) as a delivery vector in different in vitro plasmid delivery assays. Tested methods include: TP10 crosslinked to a plasmid via a peptide nucleic acid (PNA) oligomer, TP10 conjugation with polyethyleneimine (PEI), and addition of unconjugated TP10 to standard PEI transfection assay. We found that without additional DNA condensing agents, TP10 has poor transfection abilities. However, the presence of TP10 increases the transfection efficiency several folds compared to PEI alone. At as low concentrations as 0.6 nM, TP10-PNA constructs were found to enhance plasmid delivery up to 3.7-fold in Neuro-2a cells. Interestingly, the transfection efficiency was most significant at low PEI concentrations, allowing reduced PEI concentration without loss of gene delivery. No increase in cytotoxicity due to TP10 was observed and the uptake mechanism was determined to be endocytosis, as previously reported for PEI mediated transfection. In conclusion, TP10 can enhance PEI mediated transfection at relatively low concentrations and may help to develop future gene delivery systems with reduced toxicity.
authors.affiliation: Department of Neurochemistry and Neurotoxicology, Stockholm University, S-10691 Stockholm, Sweden.
authors.forename: Kalle
authors.initials: K
authors.lastname: Kilk
authors.forename: Samir
authors.initials: S
authors.lastname: El-Andaloussi
authors.forename: Peter
authors.initials: P
authors.lastname: Järver
authors.forename: Anne
authors.initials: A
authors.lastname: Meikas
authors.forename: Andres
authors.initials: A
authors.lastname: Valkna
authors.forename: Tamas
authors.initials: T
authors.lastname: Bartfai
authors.forename: Priit
authors.initials: P
authors.lastname: Kogerman
authors.forename: Madis
authors.initials: M
authors.lastname: Metsis
authors.forename: Ulo
authors.initials: U
authors.lastname: Langel
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004337:Drug Carriers / Q000737:chemistry* / Q000633:toxicity; D019004:Galanin; D018014:Gene Transfer Techniques; D006801:Humans; D020135:Peptide Nucleic Acids / Q000737:chemistry*; D010957:Plasmids / Q000008:administration & dosage*; D011094:Polyethyleneimine / Q000737:chemistry* / Q000633:toxicity; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000633:toxicity; D014162:Transfection; D014862:Wasp Venoms
chemical_list: D004337:Drug Carriers; D020135:Peptide Nucleic Acids; D011993:Recombinant Fusion Proteins; D014862:Wasp Venoms; C110068:transportan; D019004:Galanin; D011094:Polyethyleneimine