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Improving chitosan-mediated gene transfer by the introduction of intracellular buffering moieties into the chitosan backbone.

Abstract：

:Chitosan was functionalized with imidazole moieties (CHimi) with the aim of improving its buffering capacity and promoting the endosomal escape ability of chitosan-DNA complexes, ultimately increasing their transfection efficiency. 5.6%, 12.9% and 22.1% of the glucosamine residues of chitosan were substituted. Complexes with different molar ratios of primary amines to DNA phosphate anion (N/P) were prepared by a coacervation method. For an N/P>3, CHimi polymers are able to complex electrostatically with DNA and condense it into positively charged nanostructures (average size 260 nm and zeta potential +16 mV at pH 5.5). In the concentration range 2.5-100 microg ml(-1), the modified polymers had no cytotoxic effect on 293T cells. CHimi polymers with the highest degree of substitution were found to enhance beta-gal expression in 293T and HepG2 cells. Bafilomycin A1 inhibited transfection, indicating that the protonation of the imidazole groups in the endolysosome pathway favors the escape of the complexes from the endosomes, increasing the amount of transgene that can reach the cell nucleus.

journal_name

Acta Biomater

journal_title

Acta biomaterialia

authors

Moreira C,Oliveira H,Pires LR,Simões S,Barbosa MA,Pêgo AP

doi

10.1016/j.actbio.2009.04.021

subject

Has Abstract

pub_date

2009-10-01 00:00:00

pages

2995-3006

issue

8

eissn

1742-7061

issn

1878-7568

pii

S1742-7061(09)00176-7

journal_volume

5

pub_type

杂志文章

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