Characterization of Ku70(2)-NLS as bipartite nuclear localization sequence for non-viral gene delivery.

PloS One
Johannes MatschkeJoseph Rosenecker

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.

References

Aug 1, 1995·Proceedings of the National Academy of Sciences of the United States of America·O BoussifJ P Behr
Jan 18, 1994·Proceedings of the National Academy of Sciences of the United States of America·S FawellJ Barsoum
Feb 3, 1998·Nature Biotechnology·M G SebestyénJ A Wolff
Apr 29, 1998·The Journal of Biological Chemistry·H PollardD Escande
Jan 6, 1999·Proceedings of the National Academy of Sciences of the United States of America·M A ZantaJ P Behr
Jul 8, 1999·Current Opinion in Cell Biology·S A Adam
Dec 28, 1999·Annual Review of Cell and Developmental Biology·D Görlich, U Kutay
Sep 8, 2001·Molecular Therapy : the Journal of the American Society of Gene Therapy·A HonigmanE Galun
Jan 10, 2003·Journal of Radiation Research·Manabu Koike
Jan 10, 2003·The Journal of Biological Chemistry·Carsten RudolphJoseph Rosenecker
Jun 24, 2003·Biological Chemistry·Christian PlankJoseph Rosenecker
Oct 24, 2003·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Wolfgang RitterJoseph Rosenecker

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Citations

Aug 13, 2014·Journal of Peptide Science : an Official Publication of the European Peptide Society·Siegmund Reissmann
Aug 28, 2013·Journal of Controlled Release : Official Journal of the Controlled Release Society·Yuhua WangLeaf Huang
Oct 31, 2020·Molecular Pharmaceutics·Francisco E Lopez-SalasDaniel Martinez-Fong

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Methods Mentioned

BETA
transfection
nuclear translocation
PCR
electrophoresis
transfections
dynamic light scattering
FCS
protein assay
FACS

Software Mentioned

SPSS
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