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Enhanced gene delivery using disulfide-crosslinked low molecular weight polyethylenimine with listeriolysin o-polyethylenimine disulfide conjugate.

Journal of controlled release : official journal of the Controlled Release Society (2008-08-12)
Suna Choi, Kyung-Dall Lee
ABSTRACT

One of the most important requirements for non-viral gene delivery systems is the ability to mediate high levels of gene expression with low toxicity. After the DNA/vector complexes are taken up by cells through endocytosis, DNA is typically contained within the endocytic compartments and rapidly degraded due to the low pH and hydrolytic enzymes within endosomes and lysosomes, limiting its accessibility to the cytosol and ultimately to the nucleus. In this study, the endosomolytic protein listeriolysin O (LLO) from the intracellular pathogen Listeria monocytogenes was conjugated with polyethylenimine (PEI) of average molecular weight 25 kDa (PEI25) via a reversible disulfide bond (LLO-s-s-PEI), and incorporated into plasmid DNA condensed with disulfide-crosslinked low molecular weight PEI 1.8 kDa (PEI1.8). We have investigated and demonstrated that high gene transfection efficiency, which is comparable to that by the most commonly used PEI25, can be achieved by reversibly crosslinking low molecular weight PEI (PEI1.8) using disulfide bonds, with greatly reduced cytotoxicity of the PEI. The reversible incorporation of LLO into the DNA condensates of PEI, through the use of the synthesized LLO-s-s-PEI conjugate, further enhances the transfection efficiency beyond that of DNA condensates with disulfide-crosslinked PEI1.8 alone.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sodium nitrate, ACS reagent, ≥99.0%
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Sodium nitrate, ≥99.0%, suitable for plant cell culture
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Sodium nitrate, 99.995% trace metals basis
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Sodium nitrate, BioXtra, ≥99.0%
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Sodium nitrate, BioUltra, ≥99.0% (T)
Sigma-Aldrich
Sodium nitrate, anhydrous, free-flowing, Redi-Dri, ReagentPlus®, ≥99%

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