title: Non-viral gene delivery for local and controlled DNA release.
abstract: Non-viral DNA delivery systems show important advantages vs. viral systems that are usually associated with an immunological response and safety risks. In this study, disulfide cross-linked peptide-DNA condensates were investigated for local gene delivery. Two different 21 amino acid peptides were designed to have a DNA binding sequence in combination with a transglutaminase substrate site or a nuclear localization site. The peptides were used in different ratios to each other to form stable cross-linked DNA-peptide condensates with a mean diameter of 164 nm and a size distribution from 43 to 204 nm. Such aggregates showed similar stability compared to condensates formed between DNA and high molecular weight poly-L-lysine (PLL). Peptide-DNA condensates were covalently immobilized into fibrin matrices by the activity of factor XIII and were used for gene delivery in vitro. After internalization, reduction of the cross-linked peptide-DNA condensates yielded increased transfection efficiencies into different cell types cultured in 2D sandwich assays, and comparable values for HUVECs cultured in a 3D fibrin matrix, as compared to PLL-DNA condensates. Cell viability 24 h after transfection remained above 95%. The target was to develop a transfection system based on small peptides that can be covalently cross-linked into fibrin-matrices where DNA-release takes place upon cellular degradation of the matrix. This approach provides an interesting tool in non-viral gene delivery.
authors.affiliation: Institute for Biomedical Engineering and Department of Materials, ETH and University of Zurich, Moussonstrasse 18 CH-8044 Zurich, Switzerland. diana.trentin@mat.ethz.ch
authors.forename: Diana
authors.initials: D
authors.lastname: Trentin
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Hubbell
authors.forename: Heike
authors.initials: H
authors.lastname: Hall
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics; D003692:Delayed-Action Preparations / Q000008:administration & dosage* / Q000493:pharmacokinetics*; D018390:Gene Targeting / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000008:administration & dosage* / Q000138:chemical synthesis / Q000493:pharmacokinetics*; D014162:Transfection / Q000379:methods
chemical_list: D003692:Delayed-Action Preparations; D010446:Peptide Fragments; D004247:DNA