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  • The characteristics and transfection efficiency of PEI modified by biodegradable poly(beta-amino ester).

The characteristics and transfection efficiency of PEI modified by biodegradable poly(beta-amino ester).

Journal of materials science. Materials in medicine (2010-02-06)
Jong-Yuh Cherng, Yuan-Po Lee, Chao-Hsien Lin, Kui-Hsiang Chang, Wei-Yang Chang, Min-Da Shau
ABSTRACT

To improve the cytotoxicity of PEI25k and the transfection efficiency of poly(beta-amino ester) with DNA, we synthesized a poly(beta-amino ester), PEDP, bearing ester linkages in the backbone and tertiary amines in the backbone and side chain and prepared a binary mixture, PEDP-PEI25k, using physical blending meyhod. Both poly(beta-amino ester) PEDP and binary mixture PEDP-PEI25k, readily self-assembled with plasmid DNA (pCMV-beta gal) in a HEPES buffer, were characterized by dynamic light scattering. The results reveal that PEDP-PEI25k was able to self-assemble plasmid DNA into PEDP-PEI25k/DNA nano-complexes small enough to enter a cell through endocytosis. Titration studies were performed to determine the buffering capacities of PEDP and PEDP-PEI25k. The COS-7 cell viabilities in the presence of PEDP and PEDP-PEI25k were studied. At low mass ratio of PEDP/PEI25k (1/1), it is found that the transfection curve of PEDP-PEI25k/DNA bearing a maximum peak is similar to that of PEI25k/DNA. In addition, the PEDP-PEI25k/DNA complexes were able to transfect COS-7 cells in vitro with a high efficiency comparable to a well-known gene carrier PEI25k/DNA. The results indicate that binary mixture PEDP-PEI25k is an attractive cationic carrier for gene delivery and an interesting candidate for further study.

MATERIALS
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Sigma-Aldrich
HEPES buffered saline, BioUltra, for molecular biology, 2× concentrate