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Europe PubMed Central
Other literature type . 2010
Data sources: PubMed Central
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PolyPublie
Other literature type . 2010
Data sources: PolyPublie
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Molecular Therapy
Article . 2010
License: CC BY NC ND
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Intracellular Trafficking and Decondensation Kinetics of Chitosan–pDNA Polyplexes

Authors: Thibault, Marc; Nimesh, Surendra; Lavertu, Marc; Buschmann, Michael D;

Intracellular Trafficking and Decondensation Kinetics of Chitosan–pDNA Polyplexes

Abstract

The transfection efficiency (TE) of chitosan–plasmid DNA (pDNA) polyplexes can be critically modulated by the polymer degree of deacetylation (DDA) and molecular weight (MW). This study was performed to test the hypothesis that the TE dependence on chitosan MW and DDA is related to the polyplex stability, hence their intracellular decondensation/unpacking kinetics. Major barriers to nonviral gene transfer were studied by image-based quantification. Although uptake increased with increased DDA, it did not appear to be a structure-dependent process affecting TE, nor was nuclear entry. Colocalization analysis showed that all chitosans trafficked through lysosomes with similar kinetics. Fluorescent resonant energy transfer (FRET) analysis revealed a distinct relationship between TE and polyplex dissociation rate. The most efficient chitosans showed an intermediate stability and a kinetics of dissociation, which occurred in synchrony with lysosomal escape. In contrast, a rapid dissociation before lysosomal escape was found for the inefficient low DDA chitosan whereas the highly stable and inefficient complex formed by a high MW and high DDA chitosan did not dissociate even after 24 hours. This study identified that the kinetics of decondensation in relation to lysosomal escape was a most critical structure-dependent process affecting the TE of chitosan polyplexes.

Country
Canada
Related Organizations
Subjects by Vocabulary

Microsoft Academic Graph classification: Resonant inductive coupling Kinetics Dissociation (chemistry) Chitosan chemistry.chemical_compound Transfection Förster resonance energy transfer Biochemistry chemistry Acetylation Biophysics Intracellular

Keywords

Transfection, Cell Line, Drug Discovery, Fluorescence Resonance Energy Transfer, Genetics, Humans, Molecular Biology, Pharmacology, Chitosan, DNA, Flow Cytometry, Molecular Weight, Kinetics, Molecular Medicine, Original Article, Lysosomes, Plasmids

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    97
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    This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
    Top 10%
    influence
    This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    Top 10%
    impulse
    This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
    Top 10%
  • citations
    This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    97
    popularity
    This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
    Top 10%
    influence
    This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    Top 10%
    impulse
    This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
    Top 10%
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citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
97
Top 10%
Top 10%
Top 10%
Funded by
CIHR
Project
  • Funder: Canadian Institutes of Health Research (CIHR)
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