No Enhancement of Nuclear Entry by Direct Conjugation of a Nuclear Localization Signal Peptide to Linearized DNA

Tanimoto, Mitsuhide; Kamiya, Hiroyuki; Minakawa, Noriaki; Matsuda, A.; Harashima, Hideyoshi

doi:10.1021/bc034075e

Cited by 87 publications

(58 citation statements)

References 17 publications

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“…The reports indicated that the combination of G4-PAMAM dendrimer with glucocorticoid dexamethasone (PAM-Dex) facilitates the nuclear translocation of polymer/pDNA complex and increases pDNA delivery in Neuro2A cells more than two-fold and six-fold as compared to PEI and PAMAM alone, respectively 121 . The studies also demonstrated that the modification of polymers with multifunctional peptides drives the DNA from the lysosome to the nucleus 122,123 .…”

Section: Biopolymersmentioning

confidence: 99%

Physicochemical properties of polymers: An important system to overcome the cell barriers in gene transfection

et al. 2015

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Delivery of the macromolecules including DNA, miRNA, and antisense oligonucleotides is typically mediated by carriers due to the large size and negative charge. Different physical (e.g., gene gun or electroporation), and chemical (e.g., cationic polymer or lipid) vectors have been already used to improve the efficiency of gene transfer. Polymer-based DNA delivery systems have attracted special interest, in particular via intravenous injection with many intra- and extracellular barriers. The recent progress has shown that stimuli-responsive polymers entitled as multifunctional nucleic acid vehicles can act to target specific cells. These nonviral carriers are classified by the type of stimulus including reduction potential, pH, and temperature. Generally, the physicochemical characterization of DNA-polymer complexes is critical to enhance the transfection potency via protection of DNA from nuclease digestion, endosomal escape, and nuclear localization. The successful clinical applications will depend on an exact insight of barriers in gene delivery and development of carriers overcoming these barriers. Consequently, improvement of novel cationic polymers with low toxicity and effective for biomedical use has attracted a great attention in gene therapy. This article summarizes the main physicochemical and biological properties of polyplexes describing their gene transfection behavior, in vitro and in vivo. In this line, the relative efficiencies of various cationic polymers are compared.

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Section: Biopolymersmentioning

confidence: 99%

Physicochemical properties of polymers: An important system to overcome the cell barriers in gene transfection

et al. 2015

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“…In an initial trial, the NLS was directly conjugated to DNA. However, nuclear delivery efficacy is not enhanced drastically, even when 1 or 2 NLS was chemically conjugated to the linearized DNA [53,54]. Thus, modification of the pDNA with proteins (i.e.…”

Section: Control Of the Intracellular Trafficking For Pdna Deliverymentioning

confidence: 99%

“Programmed packaging” for gene delivery

Hyodo

Sakurai

Akita

et al. 2014

Journal of Controlled Release

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We report on the development of a Multifunctional Envelope-type Nano Device (MEND) based on our packaging concept "Programmed Packaging" to control not only intracellular trafficking but also the biodistribution of encapsulated compounds such as nucleic acids/proteins/peptides. Our strategy for achieving this is based on molecular mechanisms of cell biology such as endocytosis, vesicular trafficking, etc. In this review, we summarize the concept of programmed packaging and discuss some of our recent successful examples of using MENDs. Systematic evolution of ligands by exponential enrichment (SELEX) was applied as a new methodology for identifying a new ligand toward cell or mitochondria. The delivery of siRNA to tumors and the tumor vasculature was achieved using pH sensitive lipid (YSK05), which was newly designed and optimized under in vivo conditions. The efficient delivery of pDNA to immune cells such as dendritic cells has also been developed using the KALA ligand, which can be a breakthrough technology for DNA vaccine. Finally, ss-cleavable and pH-activated lipid-like surfactant (ssPalm) which is a lipid like material with pH-activatable and SS-cleavable properties is also introduced as a proof of our concept.

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“…For instance, coupling of streptavidin-NLS conjugates to biotinylated DNA enabled linear DNA fragments to enter the nuclear of digitonin-permealized cells by an active transport process. 23 Increases in gene expression has also been observed following ligation of an oligonucleotide-NLS conjugate to one or both terminal to a linear DNA molecule. 6 Conjugation of the NLS peptide to the N3-position of adenine bases in double-stranded DNA yielded no increase in the nuclear uptake of the modified DNA or transgene expression.…”

Section: Introductionmentioning

confidence: 99%

A multifunctional PEI-based cationic polyplex for enhanced systemic p53-mediated gene therapy

2006

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We recently reported a novel coupling strategy involving salicylhydroxamic acid and phenyl(di)boronic acid molecules to attach the CNGRC peptide to PEI/DNA for CD13 targeting in tumors. Here, we doubly coupled Simian Virus (SV) 40 peptide-(nuclear localization signal)) and oligonucleotidebased (DNA nuclear targeting signal) nuclear signals to the same vector using peptide nucleic acid chemistry. This vector, CNGRC/PEG/PEI/DNA-bgal/NLS/DNTS, was predominantly localized in the cell nucleus, yielding about 200-fold higher bgal gene expression in vitro, more than 20-fold increase in tumorspecific gene delivery, and a robust bgal gene expression as demonstrated in stained tumor sections. For gene therapy purposes, we further engineered a similar targeting polyplex, CNGRC/PEG/PEI/DNA-p53/NLS/DNTS, with EBV-based episomal vector for sustained p53 gene expression. A distribution of vector DNA and apoptosis in p53-containing tumors was observed, yielding a significant tumor regression and 95% animal survival after 60 days. This multicomponent vector also co-targeted tumor and tumor-associated endothelial cells but not normal cells, and had more efficient therapeutic index than each vector administered as a single modality. The use of an efficient coupling strategy without compromising the vector's integrity for DNA condensation and endosomal escape; nuclear import; tumor-specific and persistent p53 gene expression clearly provides a basis for developing a single combinatorial approach for non-viral gene therapy. Gene Therapy (2006) 13, 1512-1523.

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No Enhancement of Nuclear Entry by Direct Conjugation of a Nuclear Localization Signal Peptide to Linearized DNA

Cited by 87 publications

References 17 publications

Physicochemical properties of polymers: An important system to overcome the cell barriers in gene transfection

Physicochemical properties of polymers: An important system to overcome the cell barriers in gene transfection

“Programmed packaging” for gene delivery

A multifunctional PEI-based cationic polyplex for enhanced systemic p53-mediated gene therapy

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