原创力文档characterization of ku702–nls as bipartite nuclear localization sequence for non-viral gene delivery表征ku702-nls作为病毒性基因传递由两部分构成的核本地化序列.pdfVIP
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characterization of ku702–nls as bipartite nuclear localization sequence for non-viral gene delivery表征ku702-nls作为病毒性基因传递由两部分构成的核本地化序列
Characterization of Ku702–NLS as Bipartite Nuclear
Localization Sequence for Non-Viral Gene Delivery
1 1,2 1,3 4 1,3
Johannes Matschke , Alexander Bohla , Christof Maucksch , Rashmi Mittal , Carsten Rudolph ,
Joseph Rosenecker1*
1 Department of Pediatrics, Ludwig-Maximilians University, Munich, Germany, 2 Institute of Animal Physiology, University of Muenster, Muenster, Germany, 3 Department
of Pharmacy, Free University of Berlin, Berlin, Germany, 4 Department of Neonatology, Ludwig-Maximilians University, Munich, Germany
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 -NLS), a nuclear transport active mutant of Ku70 -NLS
2 2
(s1Ku70 -NLS) and a nuclear transport deficient mutant of Ku70 -NLS (s2Ku70 ). We examined the transfection efficiency of
2 2 2
binary Ku70 -NLS/DNA and ternary Ku70 -NLS/PEI/DNA gene vector complexes in vitro by using standard transfection
2
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