title: Characterization of Ku70(2)-NLS as bipartite nuclear localization sequence for non-viral gene delivery.
abstract: Several barriers have to be overcome in order to achieve gene expression in target cells, e.g. cellular uptake, endosomal release and translocation to the nucleus. Nuclear localization sequences (NLS) enhance gene delivery by increasing the uptake of plasmid DNA (pDNA) to the nucleus. So far, only monopartite NLS were analysed for non-viral gene delivery. In this study, we examined the characteristics of a novel bipartite NLS like construct, namely NLS Ku70. We synthesized a dimeric structure of a modified NLS from the Ku70 protein (Ku70(2)-NLS), a nuclear transport active mutant of Ku70(2)-NLS (s1Ku70(2)-NLS) and a nuclear transport deficient mutant of Ku70(2)-NLS (s2Ku70(2)). We examined the transfection efficiency of binary Ku70(2)-NLS/DNA and ternary Ku70(2)-NLS/PEI/DNA gene vector complexes in vitro by using standard transfection protocols as well as the magnetofection method. The application of Ku70(2)-NLS and s1Ku70(2)-NLS increased gene transfer efficiency in vitro and in vivo. This study shows for the first time that the use of bipartite NLS compounds alone or in combination with cationic polymers is a promising strategy to enhance the efficiency of non-viral gene transfer.
authors.affiliation: Department of Pediatrics, Ludwig-Maximilians University, Munich, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Matschke
authors.forename: Alexander
authors.initials: A
authors.lastname: Bohla
authors.forename: Christof
authors.initials: C
authors.lastname: Maucksch
authors.forename: Rashmi
authors.initials: R
authors.lastname: Mittal
authors.forename: Carsten
authors.initials: C
authors.lastname: Rudolph
authors.forename: Joseph
authors.initials: J
authors.lastname: Rosenecker
mesh_terms: D021581:Active Transport, Cell Nucleus; D034961:Antigens, Nuclear / Q000235:genetics* / Q000627:therapeutic use; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000627:therapeutic use; D018014:Gene Transfer Techniques; D000072200:Ku Autoantigen; D019913:Nuclear Localization Signals / Q000235:genetics* / Q000627:therapeutic use; D014162:Transfection
chemical_list: D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D019913:Nuclear Localization Signals; D000072200:Ku Autoantigen